Novel acyltransferases, variant thioesterases, and uses thereof

ABSTRACT

Disclosed are microalgal cells having an ablated or downregulated fatty acyl-ACP thioesterase (FATA) gene, wherein the cell is modified to express a heterologous lysophosphatidic acid acyltransferase (LPAAT) comprising an amino acid sequence that has at least 80% identity to an acyltransferase encoded by SEQ ID NO: 90, 89, 92, 93 or 95 and wherein the modified microalgal cell produces an oil with an elevated ratio of saturated-unsaturated-saturated triglycerides over trisaturated triglycerides as compared to a corresponding unmodified cell. Also disclosed are microalgal oils comprising at least 60% stearate-oleate-stearate (SOS) triglycerides, less than 5% trisaturates and wherein the fatty acid profile of the oil comprises at least 50% C18:0. Related methods of producing an oil are also disclosed.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.15/725,222, filed Oct. 4, 2017, which claims the benefit under 35 U.S.C.119(e) of U.S. Provisional Patent Application No. 62/404,667, filed Oct.5, 2016, entitled “Novel Acyltransferases, Variant Thioesterases, AndUses Thereof”, each of which is incorporated herein by reference in itsentirety for all purposes.

INCORPORATION-BY-REFERENCE OF MATERIAL SUBMITTED ELECTRONICALLY

This application includes a list of sequences, as shown at the end ofthe detailed description. The instant application contains a SequenceListing which has been submitted electronically in ASCII format and ishereby incorporated by reference in its entirety. Said ASCII copy, datedAug. 11, 2020, is named CORBP072US_SL.txt and is 606,605 bytes in size.

FIELD OF THE INVENTION

Embodiments of the present invention relate to oils/fats, fuels, foods,and oleochemicals and their production from cultures of geneticallyengineered cells. Embodiments relate to nucleic acids and proteins thatare involved in the fatty acid synthetic pathways; oils with a highcontent of triglycerides bearing fatty acyl groups upon the glycerolbackbone in particular regiospecific patterns, highly stable oils, oilswith high levels of oleic or mid-chain fatty acids, and productsproduced from such oils.

BACKGROUND OF THE INVENTION

Co-owned patent applications WO2008/151149, WO2010/063031,WO2010/063032, WO2011/150410, WO2011/150411, WO2012/061647,WO2012/061647, WO2012/106560, WO2013/158938, WO2014/120829,WO2014/151904, WO2015/051319, WO2016/007862, WO2016/014968,WO2016/044779, and WO2016/164495 relate to microbial oils and methodsfor producing those oils in host cells, including microalgae. Thesepublications also describe the use of such oils to make foods,oleochemicals, fuels and other products.

Certain enzymes of the fatty acyl-CoA elongation pathway function toextend the length of fatty acyl-CoA molecules. Elongase-complex enzymesextend fatty acyl-CoA molecules in 2 carbon additions, for examplemyristoyl-CoA to palmitoyl-CoA, stearoyl-CoA to arachidyl-CoA, oroleoyl-CoA to eicosanoyl-CoA, eicosanoyl-CoA to erucyl-CoA. In addition,elongase enzymes also extend acyl chain length in 2 carbon increments.KCS enzymes condense acyl-CoA molecules with two carbons frommalonyl-CoA to form beta-ketoacyl-CoA. KCS and elongases may showspecificity for condensing acyl substrates of particular carbon length,modification (such as hydroxylation), or degree of saturation. Forexample, the jojoba (Simmondsia chinensis) beta-ketoacyl-CoA synthasehas been demonstrated to prefer monounsaturated and saturated C18- andC20-CoA substrates to elevate production of erucic acid in transgenicplants (Lassner et al., Plant Cell, 1996, Vol 8(2), pp. 281-292),whereas specific elongase enzymes of Trypanosoma brucei show preferencefor elongating short and midchain saturated CoA substrates (Lee et al.,Cell, 2006, Vol 126(4), pp. 691-9).

The type II fatty acid biosynthetic pathway employs a series ofreactions catalyzed by soluble proteins with intermediates shuttledbetween enzymes as thioesters of acyl carrier protein (ACP). Bycontrast, the type I fatty acid biosynthetic pathway uses a single,large multifunctional polypeptide.

The oleaginous, non-photosynthetic alga, Prototheca moriformis, storescopious amounts of triacylglyceride oil under conditions when thenutritional carbon supply is in excess, but cell division is inhibiteddue to limitation of other essential nutrients. Bulk biosynthesis offatty acids with carbon chain lengths up to C18 occurs in the plastids;fatty acids are then exported to the endoplasmic reticulum where (if itoccurs) elongation past C18 and incorporation into triacylglycerides(TAGs) is believed to occur. Lipids are stored in large cytoplasmicorganelles called lipid bodies until environmental conditions change tofavor growth, whereupon they are mobilized to provide energy and carbonmolecules for anabolic metabolism.

SUMMARY OF THE INVENTION

In various aspects, the inventions disclosed herein include one or moreof the following embodiments. The embodiments can be practiced alone orin combination with each other.

Embodiment 1

This embodiment of the invention provides a recombinant vector constructor a host cell comprising nucleic acids that encode an acyltransferasethat optionally is operable to produce an altered fatty acid profile oran altered sn-2 profile in an oil produced by a host cell expressing thenucleic acids. The nucleic acids can be a nucleic acid construct or avector construct that also includes one or more regulatory elements. Theone or more regulatory elements include promoters, targeting sequences,secretion signals and other elements that control or direct theexpression of the encoded protein in the host cell. The acyltransferaseencoded by the nucleic acids have 75%, 80%, 85%, 90%, 95%, 98%, 99%, or100%, or at least 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100% identity toan acyltransferase of SEQ ID NOs: 23, 24, 25, 26, 27, 28, 29, 30, 31,32, 33, 34, 35, 36, 37, 38, 39, 40, 42, 42, 43, 44, 45, 46, 47, 48, 49,50, 52, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67,68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85,169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182,183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, or 196.The acyl transferases of this invention is a lysophosphatidic acidacyltransferase (LPAAT), glycerol phosphate acyltransferase (GPAT),diacyl glycerol acyltransferase (DGAT), lysophosphatidylcholineacyltransferase (LPCAT), or phospholipase A2 (PLA2). Theacyltransferases of the invention are shown in Table 5. In oneembodiment, the acyltransferases of the invention have acyltransferaseactivity and the amino acid sequence comprises at least 96.3%, 98%, or99% identity to an acyltransferase of clade 1 of Table 5. In anotherembodiment, the acyltransferases of the invention have acyltransferaseactivity and the amino acid sequence comprises at least 93.9%, 98%, or99% identity to an acyltransferase of clade 2 of Table 5. In oneembodiment, the acyltransferases of the invention have acyltransferaseactivity and the amino acid sequence comprises at least 86.5%, 90%, 95%,98%, or 99% identity to an acyltransferase of clade 3 of Table 5. In oneembodiment, the acyltransferases of the invention have acyltransferaseactivity and the amino acid sequence comprises at least 78.5%, 80%, 85%,90%, 95%, 98%, or 99% identity to an acyltransferase of clade 4 of Table5. In one embodiment, the recombinant vector construct of host cellcomprises nucleic acids that 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100%,or at least 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100% identity to anacyltransferase encoded by SEQ ID NOs: 19, 20, 21, 22, 88, 89, 90, 91,92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107,108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121,122, 123, 124, or 125.

Embodiment 2

This embodiment of the invention provides nucleic acids that encode anacyltransferase that when expressed produces an altered fatty acidprofile or an altered sn-2 profile in an oil produced by a host cellexpressing the nucleic acids. The nucleic acids can be a nucleic acidconstruct or a vector construct that also includes one or moreregulatory elements. The one or more regulatory elements includepromoters, targeting sequences, secretion signals and other elementsthat control or direct the expression of the encoded protein in the hostcell. The acyltransferase encoded by the nucleic acids have 75%, 80%,85%, 90%, 95%, 98%, 99%, or 100%, or at least 75%, 80%, 85%, 90%, 95%,98%, 99%, or 100% identity to an acyltransferase of SEQ ID NOs: 23, 24,25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 42, 42,43, 44, 45, 46, 47, 48, 49, 50, 52, 52, 53, 54, 55, 56, 57, 58, 59, 60,61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,79, 80, 81, 82, 83, 84, 85, 169, 170, 171, 172, 173, 174, 175, 176, 177,178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191,192, 193, 194, 195, or 196. The acyl transferases of this invention is alysophosphatidic acid acyltransferase (LPAAT), glycerol phosphateacyltransferase (GPAT), diacyl glycerol acyltransferase (DGAT),lysophosphatidylcholine acyltransferase (LPCAT), or phospholipase A2(PLA2). The acyltransferases of the invention are shown in Table 5. Inone embodiment, the acyltransferases of the invention haveacyltransferase activity and the amino acid sequence comprises at least96.3%, 98%, or 99% identity to an acyltransferase of clade 1 of Table 5.In another embodiment, the acyltransferases of the invention haveacyltransferase activity and the amino acid sequence comprises at least93.9%, 98%, or 99% identity to an acyltransferase of clade 2 of Table 5.In one embodiment, the acyltransferases of the invention haveacyltransferase activity and the amino acid sequence comprises at least86.5%, 90%, 95%, 98%, or 99% identity to an acyltransferase of clade 3of Table 5. In one embodiment, the acyltransferases of the inventionhave acyltransferase activity and the amino acid sequence comprises atleast 78.5%, 80%, 85%, 90%, 95%, 98%, or 99% identity to anacyltransferase of clade 4 of Table 5. In one embodiment, the nucleicacids comprise nucleic acids that are 75%, 80%, 85%, 90%, 95%, 98%, 99%,or 100%, or at least 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100% identityto an acyltransferase encoded by SEQ ID NOs: 19, 20, 21, 22, 88, 89, 90,91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106,107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120,121, 122, 123, 124, or 125.

Embodiment 3

This embodiment of the invention provides codon-optimized nucleic acidsthat encodes an acyltransferase operable to produce an altered fattyacid profile and/or an altered sn-2 profile in an oil produced by a hostcell expressing the nucleic acids. In one aspect, the codons areoptimized for expression in the host cell, including host cells derivedfrom plants. In another aspect, the codons are optimized for expressionin Prototheca or Chlorella. In a further aspect the codons are optimizedfor expression in Prototheca moriformis or Chlorella protothecoides. Thecodon-optimized nucleic acids can be a nucleic acid construct or avector construct that also includes one or more regulatory elements. Theone or more regulatory elements are also codon-optimized for Protothecaor Chlorella. The one or more regulatory elements include promoters,targeting sequences, secretion signals and other elements that controlor direct the expression of the encoded protein in the host cell. Theacyltransferase encoded by the codon-optimized nucleic acids have 75%,80%, 85%, 90%, 95%, 98%, 99%, or 100%, or at least 75%, 80%, 85%, 90%,95%, 98%, 99%, or 100% identity to an acyltransferase of SEQ ID NOs: 23,24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 42,42, 43, 44, 45, 46, 47, 48, 49, 50, 52, 52, 53, 54, 55, 56, 57, 58, 59,60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77,78, 79, 80, 81, 82, 83, 84, 85, 169, 170, 171, 172, 173, 174, 175, 176,177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190,191, 192, 193, 194, 195, or 196. When the codons are optimized forexpression in a host organism, at least 60%, 65%, 70%, 75%, 80%, 85%,90%, 95%, or 100% of the codons used is the most preferred codon.Alternately, at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% ofthe codons used is the first or second most preferred codon. Thecodon-optimized nucleic acids encode acyltransferases that are shown inTable 5. In one embodiment, the acyltransferases of the invention haveacyltransferase activity and the amino acid sequence comprises at least96.3%, 98%, or 99% identity to an acyltransferase of clade 1 of Table 5.In another embodiment, the acyltransferases of the invention haveacyltransferase activity and the amino acid sequence comprises at least93.9%, 98%, or 99% identity to an acyltransferase of clade 2 of Table 5.In one embodiment, the acyltransferases of the invention haveacyltransferase activity and the amino acid sequence comprises at least86.5%, 90%, 95%, 98%, or 99% identity to an acyltransferase of clade 3of Table 5. In one embodiment, the acyltransferases of the inventionhave acyltransferase activity and the amino acid sequence comprises atleast 78.5%, 80%, 85%, 90%, 95%, 98%, or 99% identity to anacyltransferase of clade 4 of Table 5. The acyltransferase encoded bythe codon-optimized nucleic acids have 75%, 80%, 85%, 90%, 95%, 98%,99%, or 100%, or at least 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100%identity to an acyltransferase of SEQ ID NOs: 23, 24, 25, 26, 27, 28,29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 42, 42, 43, 44, 45, 46,47, 48, 49, 50, 52, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64,65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82,83, 84, 85, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180,181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194,195, or 196. In one embodiment, the codon-optimizes nucleic acidscomprise nucleic acids that 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100%,or at least 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100% identity to anacyltransferase encoded by SEQ ID NOs: 19, 20, 21, 22, 88, 89, 90, 91,92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107,108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121,122, 123, 124, or 125.

Embodiment 4

In this embodiment, the invention provides host cells that areoleaginous microorganism cells or plant cells. The microorganisms of theinvention are eukaryotic microorganism. In one aspect, the host cellsare microalgae. In one embodiment, the microalgae are of the phylumChlorophyta, the class Trebouxiophytae, the order Chlorellales, or thefamily Chlorellacae. In one embodiment, the microalgae are of the genusPrototheca or Chlorella. In one embodiment, the microalgae are of thespecies Prototheca moriformis, Prototheca zopfii, Prototheca wickerhamiiPrototheca blaschkeae, Prototheca chlorelloides, Prototheca crieana,Prototheca dilamenta, Prototheca hydrocarbonea, Prototheca kruegeri,Prototheca portoricensis, Prototheca salmonis, Prototheca segbwema,Prototheca stagnorum, Prototheca trispora Prototheca ulmea, orPrototheca viscosa. Preferably, the microalga is of the speciesPrototheca moriformis. In one embodiment, the microalgae are of thespecies Chlorella autotrophica, Chlorella colonials, Chlorella lewinii,Chlorella minutissima, Chlorella pituitam, Chlorella pulchelloides,Chlorella pyrenoidosa, Chlorella rotunda, Chlorella singularis,Chlorella sorokiniana, Chlorella variabilis, or Chlorella volutis.Preferably, the microalga is of the species Chlorella protothecoides orAuxenochlorella protothecoides. The host cells express the nucleic acidsfor Embodiments relating to acyltransferases of the invention.

Embodiment 5

In this embodiment, the acyl transferase is lysophosphatidic acidacyltransferase (LPAAT), glycerol phosphate acyltransferase (GPAT),diacyl glycerol acyltransferase (DGAT), lysophosphatidylcholineacyltransferase (LPCAT), or phospholipase A2 (PLA2). In one embodiment,the acyltransferases of the invention are shown in Table 5. In oneembodiment, the acyltransferases of the invention have acyltransferaseactivity and the amino acid sequence comprises at least 96.3%, 98%, or99% identity to an acyltransferase of clade 1 of Table 5. In anotherembodiment, the acyltransferases of the invention have acyltransferaseactivity and the amino acid sequence comprises at least 93.9%, 98%, or99% identity to an acyltransferase of clade 2 of Table 5. In oneembodiment, the acyltransferases of the invention have acyltransferaseactivity and the amino acid sequence comprises at least 86.5%, 90%, 95%,98%, or 99% identity to an acyltransferase of clade 3 of Table 5. In oneembodiment, the acyltransferases of the invention have acyltransferaseactivity and the amino acid sequence comprises at least 78.5%, 80%, 85%,90%, 95%, 98%, or 99% identity to an acyltransferase of clade 4 of Table5. The acyltransferase have 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100%,or at least 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100% identity to anacyltransferase of SEQ ID NOs: 23, 24, 25, 26, 27, 28, 29, 30, 31, 32,33, 34, 35, 36, 37, 38, 39, 40, 42, 42, 43, 44, 45, 46, 47, 48, 49, 50,52, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68,69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 169,170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183,184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, or 196.

Embodiment 6

In this embodiment, nucleic acids encoding acyltransferases increasesthe production of C8:0 and/or C10:0 fatty acids or alters the sn-2profile in the host cell. The acyltransferases of the invention haveacyltransferase activity and the amino acid sequence comprises at least96.3%, 98%, or 99% identity to an acyltransferase of clade 1 of Table 5.In another embodiment, the acyltransferases of the invention haveacyltransferase activity and the amino acid sequence comprises at least93.9%, 98%, or 99% identity to an acyltransferase of clade 2 of Table 5.In one embodiment, the acyltransferases of the invention haveacyltransferase activity and the amino acid sequence comprises at least86.5%, 90%, 95%, 98%, or 99% identity to an acyltransferase of clade 3of Table 5. In one embodiment, the acyltransferases of the inventionhave acyltransferase activity and the amino acid sequence comprises atleast 78.5%, 80%, 85%, 90%, 95%, 98%, or 99% identity to anacyltransferase of clade 4 of Table 5. The C8:0 or the C10:0 content ofthe oil of the host cell is increased by 5%, 10%, 15%, 20%, 25%, 30%,35%, 40%, 45%, 50%, 55%, 60%, 70%, 80%, 90%, or higher as compared theC8:0 and/or C10:0 content of a cell oil that does not express therecombinant nucleic acids encoding the LPAATs of the invention. The sn-2profile of the oil is altered by the expression of the LPAATs of theinvention and/or the C8:0 and/or C10:0 fatty acid at the sn-2 positionis increased by 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%,60%, 70%, 80%, 90%, or higher as compared to the C8:0 and/or C10:0 fattyacid at the sn-2 position of the cell oil that does not express therecombinant nucleic acids encoding the LPAATs of the invention. Theacyltransferase encoded by the codon-optimized nucleic acids have 75%,80%, 85%, 90%, 95%, 98%, 99%, or 100%, or at least 75%, 80%, 85%, 90%,95%, 98%, 99%, or 100% identity to an acyltransferase of SEQ ID NOs: 23,24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 42,42, 43, 44, 45, 46, 47, 48, 49, 50, 52, 52, 53, 54, 55, 56, 57, 58, 59,60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77,78, 79, 80, 81, 82, 83, 84, 85, 169, 170, 171, 172, 173, 174, 175, 176,177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190,191, 192, 193, 194, 195, or 196.

Embodiment 7

This embodiment comprises nucleic acids encoding LPAATs, shown in Table5, and disclosed herein. The LPAATs encoded by the nucleic acids have75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100%, or at least 75%, 80%, 85%,90%, 95%, 98%, 99%, or 100% identity to an acyltransferase of SEQ IDNOs: 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39,40, 42, 42, 43, 44, 45, 46, 47, 48, 49, 50, 52, 52, 53, 54, 55, 56, 57,58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75,76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 169, 170, 171, 172, 173, 174,175, 176, 177, 178, 179, or 180.

Embodiment 8

In this embodiment, nucleic acids encoding GPATs of the invention have75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100%, or at least 75%, 80%, 85%,90%, 95%, 98%, 99%, or 100% identity to SEQ ID NOs: 181, 182, 183, 184,185, or 186.

Embodiment 9

In this embodiment, nucleic acids encoding DGATs of the invention have75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100%, or at least 75%, 80%, 85%,90%, 95%, 98%, 99%, or 100% identity to SEQ ID NOs: 187, or 188.

Embodiment 10

In this embodiment, nucleic acids encoding LPCATs of the invention have75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100%, or at least 75%, 80%, 85%,90%, 95%, 98%, 99%, or 100% identity to SEQ ID NOs: 189, 190, 191, or192,

Embodiment 11

This embodiment comprises nucleic acids encoding PLA2s. The PLA2sencoded by the nucleic acids have 75%, 80%, 85%, 90%, 95%, 98%, 99%, or100%, or at least 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100% identity toSEQ ID NOs: 193, 194, 195, or 196.

Embodiment 12

This embodiment is a method of cultivating a host cell expressingnucleic acids that encode the one or more acyl transferases ofembodiments 1-11

Embodiment 13

This embodiment is a method of producing an oil by cultivating hostcells that express nucleic acids that encode the one or more acyltransferases of Embodiments 1-12 and recovering the oil.

Embodiment 14

This embodiment is an oil produced by cultivating host cells thatexpress the one or more nucleic acids that encode the acyltransferasesof Examples 1-11, and recovering the oil from the host cell. When thehost cell is a microalgae, the cell oil produced by the host cell hassterols that are different than the sterols produced by a plant cell.The cell oil has a sterol profile that is different than an oil obtainedfrom a plant.

Embodiment 15

In this embodiment, a recombinant acyltransferase is provided. Therecombinant acyltransferase can be produced by a host cell. Theglycosylation of the recombinant acyl transferase is altered from theglycosylation pattern observed in the acyl transferase produced by thenon-recombinant, wild-type cell from which the gene encoding the acyltransferase was derived. In one embodiment, the recombinantacyltransferase the invention have acyltransferase activity and theamino acid sequence comprises at least 96.3%, 98%, or 99% identity to anacyltransferase of clade 1 of Table 5. In one embodiment, therecombinant acyltransferase the invention have acyltransferase activityand the amino acid sequence comprises at least 93.9%, 98%, or 99%identity to an acyltransferase of clade 2 of Table 5. In one embodiment,the acyltransferases of the invention have acyltransferase activity andthe amino acid sequence comprises at least 86.5%, 90%, 95%, 98%, or 99%identity to an acyltransferase of clade 3 of Table 5. In one embodiment,the acyltransferases of the invention have acyltransferase activity andthe amino acid sequence comprises at least 78.5%, 80%, 85%, 90%, 95%,98%, or 99% identity to an acyltransferase of clade 4 of Table 5. Theacyltransferase encoded have 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100%,or at least 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100% identity to anacyltransferase of SEQ ID NOs: 23, 24, 25, 26, 27, 28, 29, 30, 31, 32,33, 34, 35, 36, 37, 38, 39, 40, 42, 42, 43, 44, 45, 46, 47, 48, 49, 50,52, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68,69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 169,170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183,184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, or 196.

Embodiment 16

This embodiment of the invention provides a recombinant vector constructor a host cell comprising nucleic acids that encode a variant Brassicafatty acyl-ACP thioesterase that optionally is operable to produce analtered fatty acid profile in an oil produced by a host cell expressingthe nucleic acids. The nucleic acids can be a nucleic acid construct ora vector construct that also includes one or more regulatory elements.The one or more regulatory elements include promoters, targetingsequences, secretion signals and other elements that control or directthe expression of the encoded protein in the host cell. The thioesteraseencoded by the nucleic acids have 75%, 80%, 85%, 90%, 95%, 98%, 99%, or100%, or at least 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100% identity toSEQ ID NOs: 165, 166, 167, or 168 and comprise one or more of amino acidvariants D124A, D209A, D127A or D212A. In one embodiment, the BrassicaRAPA, Brassica napus or the Brassica juncea thioesterases of theinvention have fatty acyl hydrolysis activity and prefer to hydrolyzelong chain fatty acyl groups from the acyl carrier protein. In oneembodiment, the thioesterase genes, isolated from higher plants, arealtered to create variant thioesterases that have certain amino acidsthat have been altered from the wild type enzyme. Due to the alteredamino acid(s), the substrate specificity of the thioesterase is altered.The variant BnOTE enzymes increased C18:0 content by DCW, decreasedC18:1 content by DCW, and decreased C18:2 content by DCW in host cellsand the oils recovered from the host cells.

Embodiment 17

This embodiment of the invention provides a recombinant vector constructor a host cell comprising nucleic acids that encode a Garciniamangostana variant fatty acyl-ACP thioesterase (GmFATA) that optionallyis operable to produce an altered fatty acid profile in an oil producedby a host cell expressing the nucleic acids. The nucleic acids can be anucleic acid construct or a vector construct that also includes one ormore regulatory elements. The one or more regulatory elements includepromoters, targeting sequences, secretion signals and other elementsthat control or direct the expression of the encoded protein in the hostcell. The variant Garcinia thioesterase encoded by the nucleic acidshave 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100%, or at least 75%, 80%,85%, 90%, 95%, 98%, 99%, or 100% identity to SEQ ID NOs: 137, 138, 139,140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, comprise one moreof amino acid variants D variants L91F, L91K, L91S, G96A, G96T, G96V,G108A, G108V, S111A, S111V T156F, T156A, T156K, T156V, or V193A. In oneembodiment, the G. mangostana thioesterases of the invention have fattyacyl hydrolysis activity and prefer to hydrolyze long chain fatty acylgroups from the acyl carrier protein. In one embodiment, thethioesterase genes, isolated from higher plants, are altered to createvariant thioesterases that have certain amino acids that have beenaltered from the wild type enzyme. Due to the altered amino acid(s), thesubstrate specificity of the thioesterase is altered. The variant BnOTEenzymes increased C18:0 content by DCW, decreased C18:1 content by DCW,and decreased C18:2 content by DCW in host cells and the oils recoveredfrom the host cells.

Embodiment 18

This embodiment of the invention provides nucleic acids that encodevariant Brassica thioesterases or variant Garcinia thioestrases thatwhen expressed produce an altered fatty acid profile in an oil producedby a host cell expressing the nucleic acids. The nucleic acids can be anucleic acid construct or a vector construct that also includes one ormore regulatory elements. The one or more regulatory elements includepromoters, targeting sequences, secretion signals and other elementsthat control or direct the expression of the encoded protein in the hostcell. The variant Brassica thioesterases encoded by the nucleic acidshave 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100%, or at least 75%, 80%,85%, 90%, 95%, 98%, 99%, or 100% identity to SEQ ID NOs: 165, 166, 167,or 168 and comprise one or more of amino acid variants D124A, D209A,D127A or D212A. The variant variant Garcinia thioestrases encoded by thenucleic acids have 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100%, or atleast 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100% identity to SEQ ID NOs:137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150 andcomprise one or more of amino acid variants L91F, L91K, L91S, G96A,G96T, G96V, G108A, G108V, S111A, S111V T156F, T156A, T156K, T156V, orV193A.

Embodiment 19

This embodiment of the invention provides codon-optimized nucleic acidsthat encodes a variant Brassica thioesterase or a variant Garciniathioestrase operable to produce an altered fatty acid profile in an oilproduced by a host cell expressing the nucleic acids. In one aspect, thecodons are optimized for expression in the host cell, including hostcells derived from plants. In another aspect, the codons are optimizedfor expression in Prototheca or Chlorella. In a further aspect thecodons are optimized for expression in Prototheca moriformis orChlorella protothecoides. The codon-optimized nucleic acids can be anucleic acid construct or a vector construct that also includes one ormore regulatory elements. The one or more regulatory elements are alsocodon-optimized for Prototheca or Chlorella. The one or more regulatoryelements include promoters, targeting sequences, secretion signals andother elements that control or direct the expression of the encodedprotein in the host cell. The variant Brassica thioesterases encoded bythe nucleic acids have 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100%, or atleast 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100% identity to SEQ ID NOs:165, 166, 167, or 168 and comprise one or more of amino acid variantsD124A, D209A, D127A or D212A. The variant variant Garcinia thioestrasesencoded by the nucleic acids have 75%, 80%, 85%, 90%, 95%, 98%, 99%, or100%, or at least 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100% identity toSEQ ID NOs: 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148,149, or 150 and comprise one or more of amino acid variants L91F, L91K,L91S, G96A, G96T, G96V, G108A, G108V, S111A, S111V T156F, T156A, T156K,T156V, or V193A. When the codons are optimized for expression in a hostorganism, at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% ofthe codons used is the most preferred codon. Alternately, at least 60%,65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the codons used is thefirst or second most preferred codon. The codon-optimized nucleic acidsencode variant Brassica thioesterases and variant Garcinia thioestrases.In one embodiment, the variant Brassica thioesterases and variantGarcinia thioestrases of the invention have thioesterase activity.

Embodiment 20

In this embodiment, the invention provides host cells that areoleaginous microorganism cells or plant cells. The microorganisms of theinvention are eukaryotic microorganism. In one aspect, the host cellsare microalgae. In one embodiment, the microalgae are of the phylumChlorophyta, the class Trebouxiophytae, the order Chlorellales, or thefamily Chlorellacae. In one embodiment, the microalgae are of the genusPrototheca or Chlorella. In one embodiment, the microalgae are of thespecies Prototheca moriformis, Prototheca zopfii, Prototheca wickerhamiiPrototheca blaschkeae, Prototheca chlorelloides, Prototheca crieana,Prototheca dilamenta, Prototheca hydrocarbonea, Prototheca kruegeri,Prototheca portoricensis, Prototheca salmonis, Prototheca segbwema,Prototheca stagnorum, Prototheca trispora Prototheca ulmea, orPrototheca viscosa. Preferably, the microalga is of the speciesPrototheca moriformis. In one embodiment, the microalgae are of thespecies Chlorella autotrophica, Chlorella colonials, Chlorella lewinii,Chlorella minutissima, Chlorella pituitam, Chlorella pulchelloides,Chlorella pyrenoidosa, Chlorella rotunda, Chlorella singularis,Chlorella sorokiniana, Chlorella variabilis, or Chlorella volutis.Preferably, the microalga is of the species Chlorella protothecoides orAuxenochlorella protothecoides. The host cells express the nucleic acidsfor Embodiments relating to acyltransferases of the invention.

Embodiment 21

In this embodiment, the nucleic acid encoding the variant Brassicathioesterase encodes a variant thioesterase that has 75%, 80%, 85%, 90%,95%, 98%, 99%, or 100%, or at least 75%, 80%, 85%, 90%, 95%, 98%, 99%,or 100% identity to SEQ ID NOs: 165, 166, 167, or 168 and comprise oneor more of amino acid variants D124A, D209A, D127A or D212A. In anotheraspect, the nucleic acid encoding the variant Garcinia thioesteraseencodes a variant thioesterase that has 75%, 80%, 85%, 90%, 95%, 98%,99%, or 100%, or at least 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100%identity to SEQ ID NOs: 137, 138, 139, 140, 141, 142, 143, 144, 145,146, 147, 148, 149, or 150, and comprise one or more of amino acidvariants L91F, L91K, L91S, G96A, G96T, G96V, G108A, G108V, S111A, S111VT156F, T156A, T156K, T156V, or V193A.

Embodiment 22

In this embodiment, nucleic acids encoding a variant Brassicathioesterase or a variant Garcinia thioesetrase that decrease theproduction of C18:0 and/or decrease the production of C18:1 fatty acidsand/or decreases the production of C18:2 fatty acids sn-2 in the hostcell.

Embodiment 23

In this embodiment, nucleic acids encoding a variant Brassicathioesterase of the invention have SEQ ID NOs: 165, 166, 167, or 168 andcomprise one or more of amino acid variants D124A, D209A, D127A orD212A.

Embodiment 24

In this embodiment, nucleic acids encoding a variant Garciniathioesetrase of the invention have 75%, 80%, 85%, 90%, 95%, 98%, 99%, or100%, or at least 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100% identity toSEQ ID NOs: 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148,149, or 150 and comprise one or more of amino acid variants L91F, L91K,L91S, G96A, G96T, G96V, G108A, G108V, S111A, S111V T156F, T156A, T156K,T156V, or V193A.

Embodiment 25

This embodiment is a method of cultivating a host cell expressingnucleic acids that encode the one or more acyl transferases ofembodiments 16-24.

Embodiment 26

This embodiment is a method of producing an oil by cultivating hostcells that express nucleic acids that encode the one or more variantthioesterases of Embodiments 16-25 and recovering the oil.

Embodiment 27

This embodiment is an oil produced by cultivating host cells thatexpress the one or more nucleic acids that encode the varianttransferases of Examples 16-24, and recovering the oil from the hostcell. When the host cell is a microalgae, the cell oil produced by thehost cell has sterols that are different than the sterols produced by aplant cell. The cell oil has a sterol profile that is different than anoil obtained from a plant.

Embodiment 28

In this embodiment, a recombinant variant thioesterase is provided. Therecombinant variant thioesterase is produce by a host cell. Theglycosylation of the recombinant variant thioesterase is altered fromthe glycosylation pattern observed in the variant thioesterase producedby the non-recombinant, wild-type cell from which the gene encoding thevariant thioesterase was derived.

By way of example and not intended to be the only combination, theacyltransferase and/or the variant acyl-ACP thioesterrases of theinvention can be expressed in a cell in which an endogenous desaturase,KAS, and/or fatty acyl-ACP thioesterase has been ablated ordownregulated as demonstrated in the Examples. The co-expression of anacyltransferase and/or a variant acyl-ACP thioesterase concomitantlywith an invertase is an embodiment of the invention, as was demonstratedin the disclosed Examples. Additionally, the expression of anacyltansferase and/or a variant acyl-ACP thioesterase with concomitantexpression of a invertase and ablation or downregulation of adesaturase, KAS and/or fatty acyl-ACP thioesterase is an embodiment ofthe invention, as demonstrated in the disclosed Examples.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1. TAG profiles of S7815 versus the S6573 parent. TAGs in bracketsco-elute with the peak of the main TAG, but are present in traceamounts, and do not contribute significantly to the area. M=myristate(C14:0), P=palmitate (C16:0), Po=palmitoleate (C16:1), Ma=margaric(C17:0), S=stearate (C18:0), O=oleate (C18:1), L=linoleate (C18:2),Ln=linolenate (C18:3 α), A=arachidate (C20:0), B=behenate (C22:0),Lg=lignocerate (C24:0), Hx=hexacosanoate (C26:0).Sat-Sat-Sat=trisaturates. See Example 5.

FIG. 2. TAG profiles of lipids from fermentations of S7815 versus S6573.TAGs in brackets co-elute with the peak of the main TAG, but are presentin trace amounts, and do not contribute significantly to the area.M=myristate (C14:0), P=palmitate (C16:0), S=stearate (C18:0), O=oleate(C18:1), L=linoleate (C18:2), Ln=linolenate (C18:3 α), A=arachidate(C20:0), B=behenate (C22:0), Lg=lignocerate (C24:0), Hx=hexacosanoate(C26:0). Sat-Sat-Sat=trisaturates. See Example 5.

DETAILED DESCRIPTION OF THE INVENTION I. Definitions

An “allele” refers to a copy of a gene where an organism has multiplesimilar or identical gene copies, even if on the same chromosome. Anallele may encode the same or similar protein.

An “oil,” “cell oil” or “cell fat” shall mean a predominantlytriglyceride oil obtained from an organism, where the oil has notundergone blending with another natural or synthetic oil, orfractionation so as to substantially alter the fatty acid profile of thetriglyceride. In connection with an oil comprising triglycerides of aparticular regiospecificity, the cell oil or cell fat has not beensubjected to interesterification or other synthetic process to obtainthat regiospecific triglyceride profile, rather the regiospecificity isproduced naturally, by a cell or population of cells. For a cell oilproduced by a cell, the sterol profile of oil is generally determined bythe sterols produced by the cell, not by artificial reconstitution ofthe oil by adding sterols in order to mimic the cell oil. In connectionwith a cell oil or cell fat, and as used generally throughout thepresent disclosure, the terms oil, and fat are used interchangeably,except where otherwise noted. Thus, an “oil” or a “fat” can be liquid,solid, or partially solid at room temperature, depending on the makeupof the substance and other conditions. Here, the term “fractionation”means removing material from the oil in a way that changes its fattyacid profile relative to the profile produced by the organism, howeveraccomplished. The terms “oil,” “cell oil” and “cell fat” encompass suchoils obtained from an organism, where the oil has undergone minimalprocessing, including refining, bleaching, deodorized, and/or degumming,which does not substantially change its triglyceride profile. A cell oilcan also be a “noninteresterified cell oil”, which means that the celloil has not undergone a process in which fatty acids have beenredistributed in their acyl linkages to glycerol and remain essentiallyin the same configuration as when recovered from the organism.

As used herein, an oil is said to be “enriched” in one or moreparticular fatty acids if there is at least a 10% increase in the massof that fatty acid in the oil relative to the non-enriched oil. Forexample, in the case of a cell expressing a heterologous FatB genedescribed herein, the oil produced by the cell is said to be enrichedin, e.g., C8 and C16 fatty acids if the mass of these fatty acids in theoil is at least 10% greater than in oil produced by a cell of the sametype that does not express the heterologous FatB gene (e.g., wild typeoil).

“Exogenous gene” shall mean a nucleic acid that codes for the expressionof an RNA and/or protein that has been introduced into a cell (e.g. bytransformation/transfection), and is also referred to as a “transgene”.A cell comprising an exogenous gene may be referred to as a recombinantcell, into which additional exogenous gene(s) may be introduced. Theexogenous gene may be from a different species (and so heterologous), orfrom the same species (and so homologous), relative to the cell beingtransformed. Thus, an exogenous gene can include a homologous gene thatoccupies a different location in the genome of the cell or is underdifferent control, relative to the endogenous copy of the gene. Anexogenous gene may be present in more than one copy in the cell. Anexogenous gene may be maintained in a cell as an insertion into thegenome (nuclear or plastid) or as an episomal molecule.

“FADc”, also referred to as “FAD2” or “FAD” is a gene encoding adelta-12 fatty acid desaturase. “SAD” is a gene encoding a stearoyl ACPdesaturase, a delta-9 fatty acid desaturase. The desaturases desaturatesa fatty acyl chain to create a double bond. SAD converts stearic acid,C18:0 to oleic acid, C18:1 and FAD converts oleic acid, C18:1 tolinoleic acid, C18:2.

“Fatty acids” shall mean free fatty acids, fatty acid salts, or fattyacyl moieties in a glycerolipid. It will be understood that fatty acylgroups of glycerolipids can be described in terms of the carboxylic acidor anion of a carboxylic acid that is produced when the triglyceride ishydrolyzed or saponified.

“Fixed carbon source” is a molecule(s) containing carbon, typically anorganic molecule that is present at ambient temperature and pressure insolid or liquid form in a culture media that can be utilized by amicroorganism cultured therein. Accordingly, carbon dioxide is not afixed carbon source. Typical fixed carbon source include sucrose,glucose, fructose and other well-known monosaccharides, disaccharidesand polysaccharides.

“In operable linkage” is a functional linkage between two nucleic acidsequences, such a control sequence (typically a promoter) and the linkedsequence (typically a sequence that encodes a protein, also called acoding sequence). A promoter is in operable linkage with an exogenousgene if it can mediate transcription of the gene.

“Microalgae” are eukaryotic microbial organisms that contain achloroplast or other plastid, and optionally that is capable ofperforming photosynthesis, or a prokaryotic microbial organism capableof performing photosynthesis. Microalgae include obligatephotoautotrophs, which cannot metabolize a fixed carbon source asenergy, as well as heterotrophs, which can live solely off of a fixedcarbon source. Microalgae also include mixotrophic organisms that canperform photosynthesis and metabolize one or more fixed carbon source.Microalgae include unicellular organisms that separate from sister cellsshortly after cell division, such as Chlamydomonas, as well as microbessuch as, for example, volvox, which is a simple multicellularphotosynthetic microbe of two distinct cell types. Microalgae includecells such as Chlorella, Dunaliella, and Prototheca. Microalgae alsoinclude other microbial photosynthetic organisms that exhibit cell-celladhesion, such as Agmenellum, Anabaena, and Pyrobotrys. Microalgae alsoinclude obligate heterotrophic microorganisms that have lost the abilityto perform photosynthesis, such as certain dinoflagellate algae speciesand species of the genus Prototheca.

As used with respect to nucleic acids, the term “isolated” refers to anucleic acid that is free of at least one other component that istypically present with the naturally occurring nucleic acid. Thus, anaturally occurring nucleic acid is isolated if it has been purifiedaway from at least one other component that occurs naturally with thenucleic acid.

In connection with fatty acid length, “mid-chain” shall mean C8 to C16fatty acids.

In connection with a recombinant cell, the term “knockdown” refers to agene that has been partially suppressed (e.g., by about 1-95%) in termsof the production or activity of a protein encoded by the gene.Inhibitory RNA technology to down-regulate or knockdown expression of agene are well known. These techniques include dsRNA, hairpin RNA,antisense RNA, interfering RNA (RNAi) and others.

Also, in connection with a recombinant cell, the term “knockout” refersto a gene that has been completely or nearly completely (e.g., >95%)suppressed in terms of the production or activity of a protein encodedby the gene. Knockouts can be prepared by ablating the gene byhomologous recombination of a nucleic acid sequence into a codingsequence, gene deletion, mutation or other method. When homologousrecombination is performed, the nucleic acid that is inserted(“knocked-in”) can be a sequence that encodes an exogenous gene ofinterest or a sequence that does not encode for a gene of interest. Theablation by homologous recombination can be performed in one, two ormore alleles of the gene of interest.

An “oleaginous” cell is a cell capable of producing at least 20% lipidby dry cell weight, naturally or through recombinant or classical strainimprovement. An “oleaginous microbe” or “oleaginous microorganism” is amicrobe, including a microalga that is oleaginous (especially eukaryoticmicroalgae that store lipid). An oleaginous cell also encompasses a cellthat has had some or all of its lipid or other content removed, and bothlive and dead cells.

An “ordered oil” or “ordered fat” is one that forms crystals that areprimarily of a given polymorphic structure. For example, an ordered oilor ordered fat can have crystals that are greater than 50%, 60%, 70%,80%, or 90% of the 13 or 13′ polymorphic form.

In connection with a cell oil, a “profile” is the distribution ofparticular species or triglycerides or fatty acyl groups within the oil.A “fatty acid profile” is the distribution of fatty acyl groups in thetriglycerides of the oil without reference to attachment to a glycerolbackbone. Fatty acid profiles are typically determined by conversion toa fatty acid methyl ester (FAME), followed by gas chromatography (GC)analysis with flame ionization detection (FID), as in Example 1. Thefatty acid profile can be expressed as one or more percent of a fattyacid in the total fatty acid signal determined from the area under thecurve for that fatty acid. FAME-GC-FID measurement approximate weightpercentages of the fatty acids. A “sn-2 profile” is the distribution offatty acids found at the sn-2 position of the triacylglycerides in theoil. A “regiospecific profile” is the distribution of triglycerides withreference to the positioning of acyl group attachment to the glycerolbackbone without reference to stereospecificity. In other words, aregiospecific profile describes acyl group attachment at sn-1/3 vs.sn-2. Thus, in a regiospecific profile, POS (palmitate-oleate-stearate)and SOP (stearate-oleate-palmitate) are treated identically. A“stereospecific profile” describes the attachment of acyl groups atsn-1, sn-2 and sn-3. Unless otherwise indicated, triglycerides such asSOP and POS are to be considered equivalent. A “TAG profile” is thedistribution of fatty acids found in the triglycerides with reference toconnection to the glycerol backbone, but without reference to theregiospecific nature of the connections. Thus, in a TAG profile, thepercent of SSO in the oil is the sum of SSO and SOS, while in aregiospecific profile, the percent of SSO is calculated withoutinclusion of SOS species in the oil. In contrast to the weightpercentages of the FAME-GC-FID analysis, triglyceride percentages aretypically given as mole percentages; that is the percent of a given TAGmolecule in a TAG mixture.

The term “percent sequence identity,” in the context of two or moreamino acid or nucleic acid sequences, refers to two or more sequences orsubsequences that are the same or have a specified percentage of aminoacid residues or nucleotides that are the same, when compared andaligned for maximum correspondence, as measured using a sequencecomparison algorithm or by visual inspection. For sequence comparison todetermine percent nucleotide or amino acid identity, typically onesequence acts as a reference sequence, to which test sequences arecompared. When using a sequence comparison algorithm, test and referencesequences are input into a computer, subsequence coordinates aredesignated, if necessary, and sequence algorithm program parameters aredesignated. The sequence comparison algorithm then calculates thepercent sequence identity for the test sequence(s) relative to thereference sequence, based on the designated program parameters. Optimalalignment of sequences for comparison can be conducted using the NCBIBLAST software (ncbi.nlm.nih.gov/BLAST/) set to default parameters. Forexample, to compare two nucleic acid sequences, one may use blastn withthe “BLAST 2 Sequences” tool Version 2.0.12 (Apr. 21, 2000) set at thefollowing default parameters: Matrix: BLOSUM62; Reward for match: 1;Penalty for mismatch: −2; Open Gap: 5 and Extension Gap: 2 penalties;Gap x drop-off: 50; Expect: 10; Word Size: 11; Filter: on. For apairwise comparison of two amino acid sequences, one may use the “BLAST2 Sequences” tool Version 2.0.12 (Apr. 21, 2000) with blastp set, forexample, at the following default parameters: Matrix: BLOSUM62; OpenGap: 11 and Extension Gap: 1 penalties; Gap x drop-off 50; Expect: 10;Word Size: 3; Filter: on.

“Recombinant” is a cell, nucleic acid, protein or vector that has beenmodified due to the introduction of an exogenous nucleic acid or thealteration of a native nucleic acid. Thus, e.g., recombinant cells canexpress genes that are not found within the native (non-recombinant)form of the cell or express native genes differently than those genesare expressed by a non-recombinant cell. Recombinant cells can, withoutlimitation, include recombinant nucleic acids that encode for a geneproduct or for suppression elements such as mutations, knockouts,antisense, interfering RNA (RNAi), hairpin RNA or dsRNA that reduce thelevels of active gene product in a cell. A “recombinant nucleic acid” isa nucleic acid originally formed in vitro, in general, by themanipulation of nucleic acid, e.g., using polymerases, ligases,exonucleases, and endonucleases, using chemical synthesis, or otherwiseis in a form not normally found in nature. Recombinant nucleic acids maybe produced, for example, to place two or more nucleic acids in operablelinkage. Thus, an isolated nucleic acid or an expression vector formedin vitro by ligating DNA molecules that are not normally joined innature, are both considered recombinant for the purposes of thisinvention. Once a recombinant nucleic acid is made and introduced into ahost cell or organism, it may replicate using the in vivo cellularmachinery of the host cell; however, such nucleic acids, once producedrecombinantly, although subsequently replicated intracellularly, arestill considered recombinant for purposes of this invention. Similarly,a “recombinant protein” is a protein made using recombinant techniques,i.e., through the expression of a recombinant nucleic acid. Arecombinant protein will have a different pattern of glycosylation thanthe protein isolated from the wild-type organism.

The genes can be used in a variety of genetic constructs includingplasmids or other vectors for expression or recombination in a hostcell. The genes can be codon optimized for expression in a target hostcell. The proteins produced by the genes can be used in vivo or inpurified form.

For example, the gene can be prepared in an expression vector comprisingan operably linked promoter and 5′UTR. Where a plastidic cell is used asthe host, a suitably active plastid targeting peptide can be fused tothe FATB gene, as in the examples below. Generally, for the newlyidentified FATB genes, there are roughly 50 amino acids at theN-terminal that constitute a plastid transit peptide, which areresponsible for transporting the enzyme to the chloroplast. In theexamples below, this transit peptide is replaced with a 38 amino acidsequence that is effective in the Prototheca moriformis host cell fortransporting the enzyme to the plastids of those cells. Thus, theinvention contemplates deletions and fusion proteins in order tooptimize enzyme activity in a given host cell. For example, a transitpeptide from the host or related species may be used instead of that ofthe newly discovered plant genes described here.

A selectable marker gene may be included in the vector to assist inisolating a transformed cell. Examples of selectable markers useful inmicrolagae include sucrose invertase antibiotic resistance genes andother genes useful as selectable markers. The S. carlbergensis MEL1 gene(conferring the ability to grow on melibiose), A. thaliana THIC gene(conferring the ability to grow in media free of thiamine, Saccharomycessucrose invertase (conferring the ability to grow on sucrose) aredisclosed in the Examples. Other known selectable markers are useful andwithin the ambit of a skilled artisan.

The terms “triglyceride”, “triacylglyceride” and “TAG” are usedinterchangeably as is known in the art.

II. Embodiments of the Invention

Illustrative embodiments of the present invention feature oleaginouscells that produce altered fatty acid profiles and/or alteredregiospecific distribution of fatty acids in glycerolipids, and productsproduced from the cells. Examples of oleaginous cells include microbialcells having a type II fatty acid biosynthetic pathway, includingplastidic oleaginous cells such as those of oleaginous algae and, whereapplicable, oil producing cells of higher plants including but notlimited to commercial oilseed crops such as soy, corn, rapeseed/canola,cotton, flax, sunflower, safflower and peanut. Other specific examplesof cells include heterotrophic or obligate heterotrophic microalgae ofthe phylum Chlorophtya, the class Trebouxiophytae, the orderChlorellales, or the family Chlorellacae. Examples of oleaginousmicroalgae and methods of cultivation are also provided in co-ownedapplications WO2008/151149, WO2010/063031, WO2010/063032, WO2011/150410,WO2011/150411, WO2012/061647, WO2012/061647, WO2012/106560, andWO2013/158938, WO2014/120829, WO2014/151904, WO2015/051319,WO2016/007862, WO2016/014968, WO2016/044779, WO2016/164495, all of whichare incorporated by reference, including species of Chlorella andPrototheca, a genus comprising obligate heterotrophs. The oleaginouscells can be, for example, capable of producing 25%, 30%, 40%, 50%, 60%,70%, 80%, 85%, or about 90% oil by cell weight, ±5%. Optionally, theoils produced can be low in highly unsaturated fatty acids such as DHAor EPA fatty acids. For example, the oils can comprise less than 5%, 2%,or 1% DHA and/or EPA. The above-mentioned publications also disclosemethods for cultivating such cells and extracting oil, especially frommicroalgal cells; such methods are applicable to the cells disclosedherein and incorporated by reference for these teachings. Whenmicroalgal cells are used they can be cultivated autotrophically (unlessan obligate heterotroph) or in the dark using a sugar (e.g., glucose,fructose and/or sucrose) In any of the embodiments described herein, thecells can be heterotrophic cells comprising an exogenous invertase geneso as to allow the cells to produce oil from a sucrose feedstock.Alternately, or in addition, the cells can metabolize xylose fromcellulosic feedstocks. For example, the cells can be geneticallyengineered to express one or more xylose metabolism genes such as thoseencoding an active xylose transporter, a xylulose-5-phosphatetransporter, a xylose isomerase, a xylulokinase, a xylitol dehydrogenaseand a xylose reductase. See WO2012/154626, “GENETICALLY ENGINEEREDMICROORGANISMS THAT METABOLIZE XYLOSE”, published Nov. 15, 2012,including disclosure of genetically engineered Prototheca strains thatutilize xylose.

The host cells expressing the acyltransferases or the variant B. napusthioesterases or the variant G. mangostana thioesterase may, optionally,be cultivated in a bioreactor/fermenter. For example, heterotrophicoleaginous microalgal cells can be cultivated on a sugar-containingnutrient broth. Optionally, cultivation can proceed in two stages: aseed stage and a lipid-production stage. In the seed stage, the numberof cells is increased from a starter culture. Thus, the seed stage(s)typically includes a nutrient rich, nitrogen replete, media designed toencourage rapid cell division. After the seed stage(s), the cells may befed sugar under nutrient-limiting (e.g. nitrogen sparse) conditions sothat the sugar will be converted into triglycerides. As used herein,“standard lipid production conditions” are disclosed here. In oneembodiment, the culture conditions are nitrogen limiting. Sugar andother nutrients can be added during the fermentation but no additionalnitrogen is added. The cells will consume all or nearly all of thenitrogen present, but no additional nitrogen is provided. For example,the rate of cell division in the lipid-production stage can be decreasedby 50%, 80%, or more relative to the seed stage. Additionally, variationin the media between the seed stage and the lipid-production stage caninduce the recombinant cell to express different lipid-synthesis genesand thereby alter the triglycerides being produced. For example, asdiscussed below, nitrogen and/or pH sensitive promoters can be placed infront of endogenous or exogenous genes. This is especially useful whenan oil is to be produced in the lipid-production phase that does notsupport optimal growth of the cells in the seed stage.

The oleaginous cells express one or more exogenous genes encoding fattyacid biosynthesis enzymes. As a result, some embodiments feature celloils that were not obtainable from a non-plant or non-seed oil, or notobtainable at all.

The oleaginous cells, including microalgal cells, can be improved viaclassical strain improvement techniques such as UV and/or chemicalmutagenesis followed by screening or selection under environmentalconditions, including selection on a chemical or biochemical toxin. Forexample the cells can be selected on a fatty acid synthesis inhibitor, asugar metabolism inhibitor, or an herbicide. As a result of theselection, strains can be obtained with increased yield on sugar,increased oil production (e.g., as a percent of cell volume, dry weight,or liter of cell culture), or improved fatty acid or TAG profile.Co-owned application PCT/US2016/025023 filed on 31 Mar. 2016, hereinincorporated by reference, describes methods for classicallymutagenizing oleaginous cells.

The cells can be selected on one or more of 1,2-Cyclohexanedione;19-Norethindone acetate; 2,2-dichloropropionic acid;2,4,5-trichlorophenoxyacetic acid; 2,4,5-trichlorophenoxyacetic acid,methyl ester; 2,4-dichlorophenoxyacetic acid; 2,4-dichlorophenoxyaceticacid, butyl ester; 2,4-dichlorophenoxyacetic acid, isooctyl ester;2,4-dichlorophenoxyacetic acid, methyl ester; 2,4-dichlorophenoxybutyricacid; 2,4-dichlorophenoxybutyric acid, methyl ester;2,6-dichlorobenzonitrile; 2-deoxyglucose; 5-Tetradecyloxy-w-furoic acid;A-922500; acetochlor; alachlor; ametryn; amphotericin; atrazine;benfluralin; bensulide; bentazon; bromacil; bromoxynil; Cafenstrole;carbonyl cyanide m-chlorophenyl hydrazone (CCCP); carbonylcyanide-p-trifluoromethoxyphenylhydrazone (FCCP); cerulenin;chlorpropham; chlorsulfuron; clofibric acid; clopyralid; colchicine;cycloate; cyclohexamide; C75; DACTHAL (dimethyltetrachloroterephthalate); dicamba; dichloroprop((R)-2-(2,4-dichlorophenoxy)propanoic acid); Diflufenican;dihyrojasmonic acid, methyl ester; diquat; diuron; dimethylsulfoxide;Epigallocatechin gallate (EGCG); endothall; ethalfluralin; ethanol;ethofumesate; Fenoxaprop-p-ethyl; Fluazifop-p-Butyl; fluometuron;fomasefen; foramsulfuron; gibberellic acid; glufosinate ammonium;glyphosate; haloxyfop; hexazinone; imazaquin; isoxaben; Lipase inhibitorTHL ((−)-Tetrahydrolipstatin); malonic acid; MCPA(2-methyl-4-chlorophenoxyacetic acid); MCPB(4-(4-chloro-o-tolyloxy)butyric acid); mesotrione; methyldihydrojasmonate; metolachlor; metribuzin; Mildronate; molinate;naptalam; norharman; orlistat; oxadiazon; oxyfluorfen; paraquat;pendimethalin; pentachlorophenol; PF-04620110; phenethyl alcohol;phenmedipham; picloram; Platencin; Platensimycin; prometon; prometryn;pronamide; propachlor; propanil; propazine; pyrazon; Quizalofop-p-ethyl;s-ethyl dipropylthiocarbamate (EPTC); s,s,s-tributylphosphorotrithioate;salicylhydroxamic acid; sesamol; siduron; sodium methane arsenate;simazine; T-863 (DGAT inhibitor); tebuthiuron; terbacil; thiobencarb;tralkoxydim; triallate; triclopyr; triclosan; trifluralin; and vulpinicacid and others.

The oleaginous cells produce a storage oil, which is primarilytriacylglyceride and may be stored in storage bodies of the cell. A rawoil may be obtained from the cells by disrupting the cells and isolatingthe oil. The raw oil may comprise sterols produced by the cells. Patentapplications WO2008/151149, WO2010/063031, WO2010/063032, WO2011/150410,WO2011/150411, WO2012/061647, WO2012/061647, WO2012/106560,WO2013/158938, WO2014/120829, WO2014/151904, WO2015/051319,WO2016/007862, WO2016/014968, WO2016/044779, and WO2016/164495 discloseheterotrophic cultivation and oil isolation techniques for oleaginousmicroalgae. For example, oil may be obtained by providing orcultivating, drying and pressing the cells. The oils produced may berefined, bleached and deodorized (RBD) as known in the art or asdescribed in WO2010/120939. The raw or RBD oils may be used in a varietyof food, chemical, and industrial products or processes. Even after suchprocessing, the oil may retain a sterol profile characteristic of thesource. Sterol profiles of microalga and the microalgal cell oils aredisclosed below. After recovery of the oil, a valuable residual biomassremains. Uses for the residual biomass include the production of paper,plastics, absorbents, adsorbents, drilling fluids, as animal feed, forhuman nutrition, or for fertilizer.

In an embodiment of the invention nucleic acids that encode novel acyltransferases are provided. The novel acyltransferases are useful inaltering the fatty acid profile and/or altering the regiospecificprofile of an oil produced by a host cell. The nucleic acids of theinvention may contain control sequences upstream and downstream inoperable linkage with the gene of interest. These control sequencesinclude promoters, targeting sequences, untranslated sequences and othercontrol elements. Nucleic acids of the invention encode acyltransferasesthat function in type II fatty acid synthesis. The acyltransferase genesare isolated from higher plants and can be expressed in a wide varietyof host cells. The acyltransferases include lysophosphatidic acidacyltransferase (LPAAT), glycerol phosphate acyltransferase (GPAT),diacyl glycerol acyltransferase (DGAT), lysophosphatidylcholineacyltransferase (LPCAT), or phospholipase A2 (PLA2). and other lipidbiosynthetic pathway genes as discussed herein. The acyltransferases ofthe invention are shown in Table 5. In one embodiment, theacyltransferases of the invention have acyltransferase activity and theamino acid sequence comprises at least 96.3%, 98%, or 99% identity to anacyltransferase of clade 1 of Table 5. In another embodiment, theacyltransferases of the invention have acyltransferase activity and theamino acid sequence comprises at least 93.9%, 98%, or 99% identity to anacyltransferase of clade 2 of Table 5. In one embodiment, theacyltransferases of the invention have acyltransferase activity and theamino acid sequence comprises at least 86.5%, 90%, 95%, 98%, or 99%identity to an acyltransferase of clade 3 of Table 5. In one embodiment,the acyltransferases of the invention have acyltransferase activity andthe amino acid sequence comprises at least 78.5%, 80%, 85%, 90%, 95%,98%, or 99% identity to an acyltransferase of clade 4 of Table 5. Theacyltransferases when expressed increase the SOS, POP, POS, SLS, PLO,and/or PLO content DCW in host cells and the oils recovered from thehost cells. The acyltransferases when expressed in host cells decreasesthe sat-sat-sat content of the oil by DCW. The acyltransferases whenexpressed in host cells increases the sat-unsat-sat/sat-sat-sat ratio ofthe oil by DCW.

In an embodiment of the invention nucleic acids that encode variantBrassica napus thiosterases (FATA) are provided. The novel thioesterasesare useful in altering the fatty acid profile of an oil produced by ahost cell. The variant Brassica napus thiosterases prefer to hydrolyzelong chain fatty acyl groups from the acyl carrier protein. The nucleicacids of the invention may contain control sequences upstream anddownstream in operable linkage with the gene of interest. These controlsequences include promoters, targeting sequences, untranslated sequencesand other control elements. Nucleic acids of the invention encodethiosterases that function in type II fatty acid synthesis. Thethioesterase genes, isolated from higher plants, are altered to createvariant thioesterases that have certain amino acids that have beenaltered from the wild type enzyme. Due to the altered amino acid(s), thesubstrate specificity of the thioesterase is altered. The variantthioesterases can be expressed in a wide variety of host cells. Thenucleic acids encode the variant thioesterases having amino acidsequences that are 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100%, or atleast 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100% identical to SEQ IDNOs: 165, 166, 167, or 198_and comprise one or more of amino acidvariants D124A, D209A, D127A or D212A. The variant BnOTE enzymesincreased C18:0 content by DCW, decreased C18:1 content by DCW, anddecreased C18:2 content by DCW in host cells and the oils recovered fromthe host cells.

In an embodiment of the invention nucleic acids that encode variantGarcinia mangostana thiosterases (FATA) are provided. The novelthioesterases are useful in altering the fatty acid profile of an oilproduced by a host cell. The variant Garcinia mangostana thiosterasesprefer to hydrolyze long chain fatty acyl groups from the acyl carrierprotein. The nucleic acids of the invention may contain controlsequences upstream and downstream in operable linkage with the gene ofinterest. These control sequences include promoters, targetingsequences, untranslated sequences and other control elements. Nucleicacids of the invention encode thiosterases that function in type IIfatty acid synthesis. The thioesterase genes, isolated from higherplants, are altered to create variant thioesterases that have certainamino acids that have been altered from the wild type enzyme. Due to thealtered amino acid(s), the substrate specificity of the thioesterase isaltered. The variant thioesterases can be expressed in a wide variety ofhost cells. The nucleic acids encode the variant thioesterases havingamino acid sequences that are 75%, 80%, 85%, 90%, 95%, 98%, 99%, or100%, or at least 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100% identicalto SEQ ID NOs: 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147,148, 149, or 150 and comprise one or more of amino acid variants L91F,L91K, L91S, G96A, G96T, G96V, G108A, G108V, S111A, S111V T156F, T156A,T156K, T156V, or V193A. The variant GmFATA enzymes increased C18:0content by DCW, decreased C18:1 content by DCW, and decreased C18:2content by DCW in host cells and the oils recovered from the host cells.

The nucleic acids of the invention can be codon optimized for expressionin a target host cell (e.g., using the codon usage tables of Tables 1a,1b, 2a, and 2b. For example, at least 60%, 65%, 70%, 75%, 80%, 85%, 90%,95%, or 100% of the codons used can be the most preferred codonaccording to Tables 1a, 1b, 2a, and 2b. Alternately, at least 60%, 65%,70%, 75%, 80%, 85%, 90%, 95%, or 100% of the codons used can be thefirst or second most preferred codon according to Tables 1a, 1b, 2a, and2b. Preferred codons for Prototheca strains and for Chlorellaprotothecoides are shown below in Tables 1a and 1b, respectively.

TABLE 1a Preferred codon usage in Prototheca strains. Ala GCG 345 (0.36)Asn AAT 8 (0.04) GCA 66 (0.07) AAC 201 (0.96) GCT 101 (0.11) GCC 442(0.46) Pro CCG 161 (0.29) CCA 49 (0.09) Cys TGT 12 (0.10) CCT 71 (0.13)TGC 105 (0.90) CCC 267 (0.49) Asp GAT 43 (0.12) Gln CAG 226 (0.82) GAC316 (0.88) CAA 48 (0.18) Glu GAG 377 (0.96) Arg AGG 33 (0.06) GAA 14(0.04) AGA 14 (0.02) CGG 102 (0.18) Phe TTT 89 (0.29) CGA 49 (0.08) TTC216 (0.71) CGT 51 (0.09) CGC 331 (0.57) Gly GGG 92 (0.12) GGA 56 (0.07)Ser AGT 16 (0.03) GGT 76 (0.10) AGC 123 (0.22) GGC 559 (0.71) TCG 152(0.28) TCA 31 (0.06) His CAT 42 (0.21) TCT 55 (0.10) CAC 154 (0.79) TCC173 (0.31) Ile ATA 4 (0.01) Thr ACG 184 (0.38) ATT 30 (0.08) ACA 24(0.05) ATC 338 (0.91) ACT 21 (0.05) ACC 249 (0.52) Lys AAG 284 (0.98)AAA 7 (0.02) Val GTG 308 (0.50) GTA 9 (0.01) Leu TTG 26 (0.04) GTT 35(0.06) TTA 3 (0.00) GTC 262 (0.43) CTG 447 (0.61) CTA 20 (0.03) Trp TGG107 (1.00) CTT 45 (0.06) CTC 190 (0.26) Tyr TAT 10 (0.05) TAC 180 (0.95)Met ATG 191 (1.00) Stop TGA/TAG/TAA

TABLE 1b Preferred codon usage in Chlorella protothecoides. TTC (Phe)TAC (Tyr) TGC (Cys) TGA (Stop) TGG (Trp) CCC (Pro) CAC (His) CGC (Arg)CTG (Leu) CAG (Gln) ATC (Ile) ACC (Thr) GAC (Asp) TCC (Ser) ATG (Met)AAG (Lys) GCC (Ala) AAC (Asn) GGC (Gly) GTG (Val) GAG (Glu)

TABLE 2a Codon usage for Cuphea wrightii UUU F 0.48 19.5 (52) UCU S 0.2119.5 (52) UAU Y 0.45 6.4 (17) UGU C 0.41 10.5 (28) UUC F 0.52 21.3 (57)UCC S 0.26 23.6 (63) UAC Y 0.55 7.9 (21) UGC C 0.59 15.0 (40) UUA L 0.075.2 (14) UCA S 0.18 16.8 (45) UAA * 0.33 0.7 (2) UGA * 0.33 0.7 (2) UUGL 0.19 14.6 (39) UCG S 0.11 9.7 (26) UAG * 0.33 0.7 (2) UGG W 1.00 15.4(41) CUU L 0.27 21.0 (56) CCU P 0.48 21.7 (58) CAU H 0.60 11.2 (30) CGUR 0.09 5.6 (15) CUC L 0.22 17.2 (46) CCC P 0.16 7.1 (19) CAC H 0.40 7.5(20) CGC R 0.13 7.9 (21) CUA L 0.13 10.1 (27) CCA P 0.21 9.7 (26) CAA Q0.31 8.6 (23) CGA R 0.11 6.7 (18) CUG L 0.12 9.7 (26) CCG P 0.16 7.1(19) CAG Q 0.69 19.5 (52) CGG R 0.16 9.4 (25) AUU I 0.44 22.8 (61) ACU T0.33 16.8 (45) AAU N 0.66 31.4 (84) AGU S 0.18 16.1 (43) AUC I 0.29 15.4(41) ACC T 0.27 13.9 (37) AAC N 0.34 16.5 (44) AGC S 0.07 6.0 (16) AUA I0.27 13.9 (37) ACA T 0.26 13.5 (36) AAA K 0.42 21.0 (56) AGA R 0.24 14.2(38) AUG M 1.00 28.1 (75) ACG T 0.14 7.1 (19) AAG K 0.58 29.2 (78) AGG R0.27 16.1 (43) GUU V 0.28 19.8 (53) GCU A 0.35 31.4 (84) GAU D 0.63 35.9(96) GGU G 0.29 26.6 (71) GUC V 0.21 15.0 (40) GCC A 0.20 18.0 (48) GACD 0.37 21.0 (56) GGC G 0.20 18.0 (48) GUA V 0.14 10.1 (27) GCA A 0.3329.6 (79) GAA E 0.41 18.3 (49) GGA G 0.35 31.4 (84) GUG V 0.36 25.1 (67)GCG A 0.11 9.7 (26) GAG E 0.59 26.2 (70) GGG G 0.16 14.2 (38)

TABLE 2b Codon usage for Arabidopsis UUU F 0.51 21.8 (678320) UCU S 0.2825.2 (782818) UAU Y 0.52 14.6 (455089) UGU C 0.60 10.5 (327640) UUC F0.49 20.7 (642407) UCC S 0.13 11.2 (348173) UAC Y 0.48 13.7 (427132) UGCC 0.40 7.2 (222769) UUA L 0.14 12.7 (394867) UCA S 0.20 18.3 (568570)UAA * 0.36 0.9 (29405) UGA * 0.44 1.2 (36260) UUG L 0.22 20.9 (649150)UCG S 0.10 9.3 (290158) UAG * 0.20 0.5 (16417) UGG W 1.00 12.5 (388049)CUU L 0.26 24.1 (750114) CCU P 0.38 18.7 (580962) CAU H 0.61 13.8(428694) CGU R 0.17 9.0 (280392) CUC L 0.17 16.1 (500524) CCC P 0.11 5.3(165252) CAC H 0.39 8.7 (271155) CGC R 0.07 3.8 (117543) CUA L 0.11 9.9(307000) CCA P 0.33 16.1 (502101) CAA Q 0.56 19.4 (604800) CGA R 0.126.3 (195736) CUG L 0.11 9.8 (305822) CCG P 0.18 8.6 (268115) CAG Q 0.4415.2 (473809) CGG R 0.09 4.9 (151572) AUU I 0.41 21.5 (668227) ACU T0.34 17.5 (544807) AAU N 0.52 22.3 (693344) AGU S 0.16 14.0 (435738) AUCI 0.35 18.5 (576287) ACC T 0.20 10.3 (321640) AAC N 0.48 20.9 (650826)AGC S 0.13 11.3 (352568) AUA I 0.24 12.6 (391867) ACA T 0.31 15.7(487161) AAA K 0.49 30.8 (957374) AGA R 0.35 19.0 (589788) AUG M 1.0024.5 (762852) ACG T 0.15 7.7 (240652) AAG K 0.51 32.7 (1016176) AGG R0.20 11.0 (340922) GUU V 0.40 27.2 (847061) GCU A 0.43 28.3 (880808) GAUD 0.68 36.6 (1139637) GGU G 0.34 22.2 (689891) GUC V 0.19 12.8 (397008)GCC A 0.16 10.3 (321500) GAC D 0.32 17.2 (535668) GGC G 0.14 9.2(284681) GUA V 0.15 9.9 (308605) GCA A 0.27 17.5 (543180) GAA E 0.5234.3 (1068012) GGA G 0.37 24.2 (751489) GUG V 0.26 17.4 (539873) GCG A0.14 9.0 (280804) GAG E 0.48 32.2 (1002594) GGG G 0.16 10.2 (316620)

The cell oils of this invention can be distinguished from conventionalvegetable or animal triacylglycerol sources in that the sterol profilewill be indicative of the host organism as distinguishable from theconventional source. Conventional sources of oil include soy, corn,sunflower, safflower, palm, palm kernel, coconut, cottonseed, canola,rape, peanut, olive, flax, tallow, lard, cocoa, shea, mango, sal,illipe, kokum, and allanblackia.

The oils provided herein are not vegetable oils. Vegetable oils are oilsextracted from plants and plant seeds. Vegetable oils can bedistinguished from the non-plant oils provided herein on the basis oftheir oil content. A variety of methods for analyzing the oil contentcan be employed to determine the source of the oil or whetheradulteration of an oil provided herein with an oil of a different (e.g.plant) origin has occurred. The determination can be made on the basisof one or a combination of the analytical methods. These tests includebut are not limited to analysis of one or more of free fatty acids,fatty acid profile, total triacylglycerol content, diacylglycerolcontent, peroxide values, spectroscopic properties (e.g. UV absorption),sterol profile, sterol degradation products, antioxidants (e.g.tocopherols), pigments (e.g. chlorophyll), d13C values and sensoryanalysis (e.g. taste, odor, and mouth feel). Many such tests have beenstandardized for commercial oils such as the Codex Alimentariusstandards for edible fats and oils.

Sterol profile analysis is a particularly well-known method fordetermining the biological source of organic matter. Campesterol,b-sitosterol, and stigamsterol are common plant sterols, withb-sitosterol being a principle plant sterol. For example, b-sitosterolwas found to be in greatest abundance in an analysis of certain seedoils, approximately 64% in corn, 29% in rapeseed, 64% in sunflower, 74%in cottonseed, 26% in soybean, and 79% in olive oil (Gul et al. J. Celland Molecular Biology 5:71-79, 2006).

The sterol profile of a microalgal oil is distinct from the sterolprofile of oils obtained from higher plants or animals. Oil isolatedfrom Prototheca moriformis strain UTEX1435 were separately clarified(CL), refined and bleached (RB), or refined, bleached and deodorized(RBD) and were tested for sterol content according to the proceduredescribed in JAOCS vol. 60, no. 8, August 1983. Results of the analysisare shown Table 3 below (units in mg/100 g):

TABLE 3 (units in mg/100 g) Refined, Refined & bleached, & Sterol CrudeClarified bleached deodorized 1 Ergosterol 384   398   293   302   (56%)  (55%)  (50%)  (50%) 2 5,22-cholestadien-24- 14.6 18.8 14   15.2methyl-3-ol (2.1%) (2.6%) (2.4%) (2.5%) (Brassicasterol) 324-methylcholest-5- 10.7 11.9 10.9 10.8 en-3-ol (Campesterol or (1.6%)(1.6%) (1.8%) (1.8%) 22,23- dihydrobrassicasterol) 45,22-cholestadien-24- 57.7 59.2 46.8 49.9 ethyl-3-ol (Stigmasterol(8.4%) (8.2%) (7.9%) (8.3%) or poriferasterol) 5 24-ethylcholest-5-en- 9.64  9.92  9.26 10.2 3-ol (β-Sitosterol or (1.4%) (1.4%) (1.6%) (1.7%)clionasterol) 6 Other sterols 209   221   216   213   Total sterols685.64 718.82 589.96 601.1 

These results show three striking features. First, ergosterol was foundto be the most abundant of all the sterols, accounting for about 50% ormore of the total sterols. The amount of ergosterol is greater than thatof campesterol, β-sitosterol, and stigmasterol combined. Ergosterol issteroid commonly found in fungus and not commonly found in plants, andits presence particularly in significant amounts serves as a usefulmarker for non-plant oils. Secondly, the oil was found to containbrassicasterol. With the exception of rapeseed oil, brassicasterol isnot commonly found in plant based oils. Thirdly, less than 2%β-sitosterol was found to be present. β-sitosterol is a prominent plantsterol not commonly found in microalgae, and its presence particularlyin significant amounts serves as a useful marker for oils of plantorigin. In summary, Prototheca moriformis strain UTEX1435 has been foundto contain both significant amounts of ergosterol and only trace amountsof β-sitosterol as a percentage of total sterol content. Accordingly,the ratio of ergosterol:β-sitosterol or in combination with the presenceof brassicasterol can be used to distinguish this oil from plant oils.

In some embodiments, the oil content of an oil provided herein contains,as a percentage of total sterols, less than 20%, 15%, 10%, 5%, 4%, 3%,2%, or 1% β-sitosterol. In other embodiments the oil is free fromβ-sitosterol.

In some embodiments, the oil is free from one or more of β-sitosterol,campesterol, or stigmasterol. In some embodiments the oil is free fromβ-sitosterol, campesterol, and stigmasterol. In some embodiments the oilis free from campesterol. In some embodiments the oil is free fromstigmasterol.

In some embodiments, the oil content of an oil provided hereincomprises, as a percentage of total sterols, less than 20%, 15%, 10%,5%, 4%, 3%, 2%, or 1% 24-ethylcholest-5-en-3-ol. In some embodiments,the 24-ethylcholest-5-en-3-ol is clionasterol. In some embodiments, theoil content of an oil provided herein comprises, as a percentage oftotal sterols, at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or 10%clionasterol.

In some embodiments, the oil content of an oil provided herein contains,as a percentage of total sterols, less than 20%, 15%, 10%, 5%, 4%, 3%,2%, or 1% 24-methylcholest-5-en-3-ol. In some embodiments, the24-methylcholest-5-en-3-ol is 22, 23-dihydrobrassicasterol. In someembodiments, the oil content of an oil provided herein comprises, as apercentage of total sterols, at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%,9%, or 10% 22,23-dihydrobrassicasterol.

In some embodiments, the oil content of an oil provided herein contains,as a percentage of total sterols, less than 20%, 15%, 10%, 5%, 4%, 3%,2%, or 1% 5,22-cholestadien-24-ethyl-3-ol. In some embodiments, the 5,22-cholestadien-24-ethyl-3-ol is poriferasterol. In some embodiments,the oil content of an oil provided herein comprises, as a percentage oftotal sterols, at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or 10%poriferasterol.

In some embodiments, the oil content of an oil provided herein containsergosterol or brassicasterol or a combination of the two. In someembodiments, the oil content contains, as a percentage of total sterols,at least 5%, 10%, 20%, 25%, 35%, 40%, 45%, 50%, 55%, 60%, or 65%ergosterol. In some embodiments, the oil content contains, as apercentage of total sterols, at least 25% ergosterol. In someembodiments, the oil content contains, as a percentage of total sterols,at least 40% ergosterol. In some embodiments, the oil content contains,as a percentage of total sterols, at least 5%, 10%, 20%, 25%, 35%, 40%,45%, 50%, 55%, 60%, or 65% of a combination of ergosterol andbrassicasterol.

In some embodiments, the oil content contains, as a percentage of totalsterols, at least 1%, 2%, 3%, 4%, or 5% brassicasterol. In someembodiments, the oil content contains, as a percentage of total sterolsless than 10%, 9%, 8%, 7%, 6%, or 5% brassicasterol.

In some embodiments the ratio of ergosterol to brassicasterol is atleast 5:1, 10:1, 15:1, or 20:1.

In some embodiments, the oil content contains, as a percentage of totalsterols, at least 5%, 10%, 20%, 25%, 35%, 40%, 45%, 50%, 55%, 60%, or65% ergosterol and less than 20%, 15%, 10%, 5%, 4%, 3%, 2%, or 1%β-sitosterol. In some embodiments, the oil content contains, as apercentage of total sterols, at least 25% ergosterol and less than 5%β-sitosterol. In some embodiments, the oil content further comprisesbrassicasterol.

Sterols contain from 27 to 29 carbon atoms (C27 to C29) and are found inall eukaryotes. Animals exclusively make C27 sterols as they lack theability to further modify the C27 sterols to produce C28 and C29sterols. Plants however are able to synthesize C28 and C29 sterols, andC28/C29 plant sterols are often referred to as phytosterols. The sterolprofile of a given plant is high in C29 sterols, and the primary sterolsin plants are typically the C29 sterols b-sitosterol and stigmasterol.In contrast, the sterol profiles of non-plant organisms contain greaterpercentages of C27 and C28 sterols. For example the sterols in fungi andin many microalgae are principally C28 sterols. The sterol profile andparticularly the striking predominance of C29 sterols over C28 sterolsin plants has been exploited for determining the proportion of plant andmarine matter in soil samples (Huang, Wen-Yen, Meinschein W. G.,“Sterols as ecological indicators”; Geochimica et Cosmochimia Acta. Vol43. pp 739-745).

In some embodiments the primary sterols in the microalgal oils providedherein are sterols other than b-sitosterol and stigmasterol. In someembodiments of the microalgal oils, C29 sterols make up less than 50%,40%, 30%, 20%, 10%, or 5% by weight of the total sterol content.

In some embodiments the microalgal oils provided herein contain C28sterols in excess of C29 sterols. In some embodiments of the microalgaloils, C28 sterols make up greater than 50%, 60%, 70%, 80%, 90%, or 95%by weight of the total sterol content. In some embodiments the C28sterol is ergosterol. In some embodiments the C28 sterol isbrassicasterol.

Where a fatty acid profile of a triglyceride (also referred to as a“triacylglyceride” or “TAG”) cell oil is given here, it will beunderstood that this refers to a nonfractionated sample of the storageoil extracted from the cell analyzed under conditions in whichphospholipids have been removed or with an analysis method that issubstantially insensitive to the fatty acids of the phospholipids (e.g.using chromatography and mass spectrometry). The oil may be subjected toan RBD process to remove phospholipids, free fatty acids and odors yethave only minor or negligible changes to the fatty acid profile of thetriglycerides in the oil. Because the cells are oleaginous, in somecases the storage oil will constitute the bulk of all the TAGs in thecell. Examples 1 and 2 below give analytical methods for determining TAGfatty acid composition and regiospecific structure.

Broadly categorized, certain embodiments of the invention include (i)recombinant oleaginous cells that comprise an ablation of one or two orall alleles of an endogenous polynucleotide, including polynucleotidesencoding lysophosphatidic acid acyltransferase (LPAAT) or (ii) cellsthat produce oils having low concentrations of polyunsaturated fattyacids, including cells that are auxotrophic for unsaturated fatty acids;(iii) cells producing oils having high concentrations of particularfatty acids due to expression of one or more exogenous genes encodingenzymes that transfer fatty acids to glycerol or a glycerol ester; (iv)cells producing regiospecific oils, (v) genetic constructs or cellsencoding a an LPAAT, a lysophosphatidylcholine acyltransferase (LPCAT),a phosphatidylcholine diacylglycerol cholinephosphotransferase (PDCT),diacylglycerol cholinephosphotransferase (DAG-CPT) or fatty acylelongase (FAE), (vi) cells producing low levels of saturated fatty acidsand/or high levels of C18:1, C18:2, C18:3, C20:1 or C22:1, (vii) andother inventions related to producing cell oils with altered profiles.The embodiments also encompass the oils made by such cells, the residualbiomass from such cells after oil extraction, oleochemicals, fuels andfood products made from the oils and methods of cultivating the cells.

In any of the embodiments below, the cells used are optionally cellshaving a type II fatty acid biosynthetic pathway such as plant cells,yeast cells, microalgal cells including heterotrophic or obligateheterotrophic microalgal cells, including cells classified asChlorophyta, Trebouxiophyceae, Chlorellales, Chlorellaceae, orChlorophyceae, or cells engineered to have a type II fatty acidbiosynthetic pathway using the tools of synthetic biology (i.e.,transplanting the genetic machinery for a type II fatty acidbiosynthesis into an organism lacking such a pathway). Use of a hostcell with a type II pathway avoids the potential for non-interactionbetween an exogenous acyl-ACP thioesterase or other ACP-binding enzymeand the multienzyme complex of type I cellular machinery. In specificembodiments, the cell is of the species Prototheca moriformis,Prototheca krugani, Prototheca stagnora or Prototheca zopfii or has a23S rRNA sequence with at least 65, 70, 75, 80, 85, 90 or 95% nucleotideidentity SEQ ID NO: 25. By cultivating in the dark or using an obligateheterotroph, the cell oil produced can be low in chlorophyll or othercolorants. For example, the cell oil can have less than 100, 50, 10, 5,1, 0.0.5 ppm of chlorophyll without substantial purification.

The stable carbon isotope value δ13C is an expression of the ratio of¹³C/¹²C relative to a standard (e.g. PDB, carbonite of fossil skeletonof Belemnite americana from Peedee formation of South Carolina). Thestable carbon isotope value δ13C (‰) of the oils can be related to theδ13C value of the feedstock used. In some embodiments the oils arederived from oleaginous organisms heterotrophically grown on sugarderived from a C4 plant such as corn or sugarcane. In some embodimentsthe δ13C (‰) of the oil is from −10 to −17‰ or from −13 to −16‰.

In specific embodiments and examples discussed below, one or more fattyacid synthesis genes (e.g., encoding an acyl-ACP thioesterase, aketo-acyl ACP synthase, an LPAAT, an LPCAT, a PDCT, a DAG-CPT, an FAE astearoyl ACP desaturase, or others described herein) is incorporatedinto a microalga. It has been found that for certain microalga, a plantfatty acid synthesis gene product is functional in the absence of thecorresponding plant acyl carrier protein (ACP), even when the geneproduct is an enzyme, such as an acyl-ACP thioesterase, that requiresbinding of ACP to function. Thus, optionally, the microalgal cells canutilize such genes to make a desired oil without co-expression of theplant ACP gene.

For the various embodiments of recombinant cells comprising exogenousgenes or combinations of genes, it is contemplated that substitution ofthose genes with genes having 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%,94%, 95%, 96%, 97%, 98%, or 99% or 100% nucleic acid sequence identitycan give similar results, as can substitution of genes encoding proteinshaving 60%, 70%, 80%, 85%, 90%, 91% 92%, 93%, 94%, 95%, 95.5%, 96%,96.5%, 97%, 97.5%, 98%, 98.5%, 99% or 100% amino acid sequence identity.Nucleic acids encoding the acyltransferases encode acyltransferases thathave 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100%, or atleast 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100% aminoacid sequence identity to the acyltransferase disclosed in clade 1,clade 2, clade 3 or clade 4 of Table 5. Likewise, for novel regulatoryelements, it is contemplated that substitution of those nucleic acidswith nucleic acids having 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%,95%, 96%, 97%, 98%, 99% or 100% nucleic acid can be efficacious. In thevarious embodiments, it will be understood that sequences that are notnecessary for function (e.g. FLAG® tags or inserted restriction sites)can often be omitted in use or ignored in comparing genes, proteins andvariants.

The novel genes and gene combinations reported here can be used inhigher plants using techniques that are well known in the art. Forexample, the use of exogenous lipid metabolism genes in higher plants isdescribed in U.S. Pat. Nos. 6,028,247; 5,850,022; 5,639,790; 5,455,167;5,512,482; and 5,298,421 disclose higher plants with exogenous acyl-ACPthioesterases. WO2009129582 and WO1995027791 disclose cloning of LPAATin plants. FAD2 ablation and/or down regulation in higher plants istaught in WO 2013112578, and WO2008/006171. SAD ablation and/or downregulation in higher plants is taught in WO 2013112578, and WO2008006171.

The expression of the novel acyltransferases is shown in Examples 4, 5,6 and 7. The expression of Cuphea paucipetala or Cuphea ignea LPATsmarkedly increased the C8:0 and C10:0 fraction of the cell oil.Additionally, the expression of Cuphea paucipetala or Cuphea igneaLPAATs markedly increased the incorporation of C8:0 and C10:0 fattyacids in the sn-2 position of the TAG. This is disclosed in Example 4.

The expression of LPAT genes in host cells increased C18:2 levels andelevated the sat-unsat-sat/sat-sat-sat, (e.g., SOS/SSS) ratio of thecell oil. For example, the expression of Theobroma cacoa LPAT2 drivesthe transfer of unsaturated fatty acids toward the sn-2 position andreduces the incorporation of saturated fatty acids at sn-2.

The novel LPAATs, GPATs, DGATs, LPCATs, and PLA2 with specificity formid-chain fatty acids are disclosed. In Example 7, expression of LPAATsand DGATs are disclosed.

When an acyltransferase of the invention is expressed in a host cell,one or more additional exogenous genes can concomitantly be expressed.An embodiment of this invention provides host cells that express arecombinant acyltransferase and concomitantly express one or moreadditional recombinant genes. The one or more additional genes includeinvertase, fatty acyl-ACP thioesterase (FATA, FATB), melibiase, ketoacylsynthase (KASI, KASII, KASIII, KASIV), antibiotic selective markers,tags such as FLAG, and THIC. In Examples 4, 5, 6, and 7, theco-expression of nucleic acids that encode LPAATs co-expressed with oneor more exogenous genes that encode invertase, fatty acyl-ACPthioesterase, melibiase, ketoacyl synthase, THIC are disclosed.

When an acyltransferase of the invention is expressed in a host cell, anendogenous gene of the host call can concomitantly be ablated ordownregulated, thereby eliminating or decreasing the expression of thegene of the host cell. This can be accomplished by using homologousrecombination techniques or other RNA inhibitory technologies. Theablated or downregulated gene can be any gene in the host cell. Theablated or downregulated endogenous gene can be stearoyl ACP desaturase,fatty acyl desaturase, fatty acyl-ACP thioesterase (FATA or FATB),ketoacyl synthase (KASI, KASII, KASIII or KAS IV), or an acyltransferase(LPAAT, DGAT, GPAT, LPCAT). When an endogenous is ablated, one, two ormore alleles of the endogenous can be ablated. In Example 5, theexpression of a Brassica LPAAT, while concomitantly ablating anendogenous stearoyl ACP desaturase is disclosed. In Example 6, LPAATs,GPATs, DGATs, LPCATs and PLA2s with specificity for mid-chain fattyacids were expressed, while ablating a gene encoding stearoyl ACPdesaturase. In Example 7 the down regulation of an endogenous FAD2 and ahairpin RNA is disclosed. In co-owned PCT/US2016/026265, applicantsdisclosed concomitant ablation of an endogenous LPAAT and expression ofan exogenous LPAAT.

In one embodiment, the expression of the acyl transferases alters thefatty acid profile and/or the sn-2 profile of the oil produced by thehost organism. The fatty acid profiles and the sn-2 profiles that resultfrom the expression of various acyltransferases are disclosed in Tables6, 7, 10, 11, 12, 13, 16, 17, 18, 19, 20, 22, 23, and 24. The inventionprovides host cells with altered fatty acid profiles and altered sn-2profiles according to Tables 6, 7, 10, 11, 12, 13, 16, 17, 18, 19, 20,22, 23, and 24.

As described in PCT/US2016/026265, co-owned by applicant, transcriptprofiling was used to discover promoters that modulate expression inresponse to low nitrogen conditions. The promoters are useful toselectively express various genes and to alter the fatty acidcomposition of microbial oils. In accordance with an embodiment, thereare non-natural constructs comprising a heterologous promoter and agene, wherein the promoter comprises at least 60%, 65%, 70%, 75%, 80%,85%, 90%, 95%, or 100% sequence identity to any of the promoters of SEQID NOs: 1-18 and the gene is differentially expressed under low vs. highnitrogen conditions. In particular, the Prototheca moriformis AMT02 (SEQID NO: 18) and AMT03 promoter (SEQ ID NO: 18) are useful promoters forcontrolling the expression of an exogenous gene. For example, thepromoters can be placed in front of a FAD2 gene in a linoleic acidauxotroph to produce an oil with less than 5, 4, 3, 2, or 1% linoleicacid after culturing first under high nitrogen conditions, then nextculturing under low nitrogen conditions. Additional promoters, inparticulare Prototheca and Chlorella promoters are described in thesequences and descriptions in this application. For example, thePrototheca HXT1, SAD, LDH1 and other Prototheca promoters are describedin Examples 6, 7, 8, and 9. Additionally, the Chlorella SAD, ACT andother Chlorella promoters are described in Examples 6, 7, 8, and 9.

In embodiments of the present invention, oleaginous cells expressing oneor more of the genes encoding acyltransferases and/or variant FATA canproduce an oil with at least 20, 40, 60 or 70% of C8, C10, C12, C14,C16, or C18 fatty acids.

The invention also provides host cells expressing one or more of thegenes encoding acyltransferases and/or variant FATA can produce an oilenriched is oils that are sat-unsat-sat. Oils of this type include SOS,POP, POS, SLS, PLO, PLO. The sat-unsat-sat oils comprise at least 30%,40%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95% of the cell oil bydry cell weight.

The invention also provides host cells expressing one or more of thegenes encoding acyltransferases and/or variant FATA can produce an oilthat is decreased in tri-saturated oils, sat-sat-sat. Oils of this typeinclude PPP, PSS, PPS, SSS, SPS, and PSP. The sat-sat-sat oils compriseless than 50%, 40%, 30%, 20%, 15%, 10%, 8%, 6%, 5%, 4%, 3%, 2%, or 1% ofthe cell oil by molar fraction or dry cell weight.

The host cells of the invention can produce 25%, 30%, 40%, 50%, 60%,70%, 80%, 85%, or about 90% oil by cell weight, ±5%. Optionally, theoils produced can be low in DHA or EPA fatty acids. For example, theoils can comprise less than 5%, 2%, or 1% DHA and/or EPA.

In other embodiments of the invention, there is a process for producingan oil, triglyceride, fatty acid, or derivative of any of these,comprising transforming a cell with any of the nucleic acids discussedherein. In another embodiment, the transformed cell is cultivated toproduce an oil and, optionally, the oil is extracted. Oil extracted inthis way can be used to produce food, oleochemicals or other products.

The oils discussed above alone or in combination are useful in theproduction of foods, fuels and chemicals (including plastics, foams,films, etc). The oils, triglycerides, fatty acids from the oils may besubjected to C—H activation, hydroamino methylation,methoxy-carbonation, ozonolysis, enzymatic transformations, epoxidation,methylation, dimerization, thiolation, metathesis, hydro-alkylation,lactonization, or other chemical processes.

After extracting the oil, a residual biomass may be left, which may haveuse as a fuel, as an animal feed, or as an ingredient in paper, plastic,or other product. For example, residual biomass from heterotrophic algaecan be used in such products.

EXAMPLES Example 1: Fatty Acid Analysis by Fatty Acid Methyl EsterDetection

Lipid samples were prepared from dried biomass. 20-40 mg of driedbiomass was resuspended in 2 mL of 5% H₂SO₄ in MeOH, and 200 ul oftoluene containing an appropriate amount of a suitable internal standard(C19:0) was added. The mixture was sonicated briefly to disperse thebiomass, then heated at 70-75° C. for 3.5 hours. 2 mL of heptane wasadded to extract the fatty acid methyl esters, followed by addition of 2mL of 6% K₂CO₃ (aq) to neutralize the acid. The mixture was agitatedvigorously, and a portion of the upper layer was transferred to a vialcontaining Na₂SO₄ (anhydrous) for gas chromatography analysis usingstandard FAME GC/FID (fatty acid methyl ester gas chromatography flameionization detection) methods. Fatty acid profiles reported below weredetermined by this method.

Example 2: Analysis of Regiospecific Profile

LC/MS TAG distribution analyses were carried out using a Shimadzu Nexeraultra high performance liquid chromatography system that included aSIL-30AC autosampler, two LC-30AD pumps, a DGU-20A5 in-line degasser,and a CTO-20A column oven, coupled to a Shimadzu LCMS 8030 triplequadrupole mass spectrometer equipped with an APCI source. Data wasacquired using a Q3 scan of m/z 350-1050 at a scan speed of 1428 u/secin positive ion mode with the CID gas (argon) pressure set to 230 kPa.The APCI, desolvation line, and heat block temperatures were set to 300,250, and 200° C., respectively, the flow rates of the nebulizing anddrying gases were 3.0 L/min and 5.0 L/min, respectively, and theinterface voltage was 4500 V. Oil samples were dissolved indichloromethane-methanol (1:1) to a concentration of 5 mg/mL, and 0.8 μLof sample was injected onto Shimadzu Shim-pack XR-ODS III (2.2 μm,2.0×200 mm) maintained at 30° C. A linear gradient from 30%dichloromethane-2-propanol (1:1)/acetonitrile to 51%dichloromethane-2-propanol (1:1)/acetonitrile over 27 minutes at 0.48mL/min was used for chromatographic separations.

Example 3: Cultivation of Microalgae Standard Lipid ProductionConditions:

Cells scraped from a source plate with toothpicks were used to inoculatepre-seed cultures of 0.5 mL EB03, 0.5% glucose, 1×DAS2 cultures in96-well blocks. Pre-seed cultures were grown for 70-75 h at 28° C., 900rpm in a Multitron shaker. 40 μl of pre-seed cultures were used toinoculate seed cultures of 0.46 mL H29, 4% glucose, 25 mM citrate pH 5or 100 mM PIPES pH 7.3, 1×DAS2 (8% inoculum), and grown for 24-28 h at28° C., 900 rpm in a Multitron shaker. 40 μL of seed cultures were usedto inoculate lipid production cultures of 0.46 mL H43, 6% glucose, 25 mMcitrate pH 5, 1×DAS2 (8% inoculum), and grown for 70-75 h at 28° C., 900rpm in a Multitron shaker. Fatty acid profiles and lipid titer analyseswere performed as disclosed in Examples 1 and 2.

50 mL Shake Flask Format

Cells scraped from a source plate with inoculation loops, or cellcultures from cryovials were used to inoculate pre-seed cultures of 10mL EB03, 0.5% glucose, 1×DAS2 cultures in 50 mL bioreactor tubes.Pre-seed cultures were grown for 70-75 h at 28° C., 200 rpm in a Kuhnershaker. 0.8 mL of pre-seed cultures were used to inoculate seed culturesof 10 mL H29, 4% glucose, 25 mM citrate pH 5 or 100 mM PIPES pH 7.3,1×DAS2 (8% inoculum), and grown for 24-28 h at 28° C., 200 rpm in aKuhner shaker. 100 μL of seed cultures were used to inoculate lipidproduction cultures of 49.9 mL H43, 6% glucose, 25 mM citrate pH 5 or100 mM PIPES pH 7.3, 1×DAS2 (0.2% inoculum), and grown for 118-122 h at28° C., 200 rpm in a Kuhner shaker. Fatty acid profiles and lipid titeranalyses were performed as disclosed in Examples 1 and 2.

EB03

Dry chemicals Component Concentration (g/L) K2HPO4 3 Sodium PhosphateDibasic Heptahydrate 5.66 (Na2HPO4 7H2O) citric acid monohydrate 1.2ammonium sulfate 1 MgSO4 7H2O 0.23 CaCl2 2H2O 0.03 Stock solutionsComponent Concentration (mL/L) 100X C-Trace (3) 10 Antifoam Sigma 2040.225

H29

Dry chemicals Final Component Concentration (g/L) K₂HPO₄ (Potassiumphosphate 0.25 dibasic anhydrous) NaH₂PO₄ (Sodium phosphate 0.18monobasic) MgSO₄•7H₂O (Magnesium 0.24 sulfate heptahydrate) Citric acidmonohydrate 0.25 Stock solutions Component Concentration (mL/L) 0.017Mstock CaCl₂•2H₂O 10 0.151M (NH₄)₂SO₄ 52.2 100X C-Trace (2) 10 AntifoamSigma 204 0.225

H43

Dry chemicals Final Component Concentration (g/L) K2HPO4 0.25 NaH2PO40.18 MgSO4 7H2O 0.24 Citric acid H2O 0.25 Stock solutions ComponentConcentration (mL/L) 0.017M stock CaCl2 2H2O 10 100X C-Trace (2) 10Antifoam Sigma 204 0.225 0.151M (NH4)2SO4 12.5

1000×DAS2

Dry chemicals Final Component Concentration (g/L) Thiamine-HCl 0.67d-Biotin 0.010 Cyanocobalimin (vit B-12) 0.008 Calcium Pantothenate 0.02PABA (p-aminobenzoic acid) 0.04

100×C-Trace(2)

Dry chemicals Final Component Concentration (g/L) CuSO4—5H2O 0.011CoC12—6H2O 0.081 H3BO3 0.33 ZnSO4—7H2O 1.4 MnSO4—H2O 0.81 Na2MoO4—2H2O0.039 FeSO4—7H2O 0.11 NiCl2—6H2O 0.013 Citric Acid Monohydrate 3.0

100×C-Trace (3)

Dry chemicals Final Component Concentration (g/L) CuSO4—5H2O 0.011 H3BO30.33 ZnSO4—7H2O 1.4 MnSO4—H2O 0.81 Na2MoO4—2H2O 0.039 FeSO4—7H2O 0.11NiCl2—6H2O 0.013 Citric Acid Monohydrate 3.0

Example 4: Identification of Novel LPAAT Genes from SequencedTranscriptomes and Engineering Sn-2 Tag Regiospecificity in UTEX1435 byExpression of Heterologous LPAAT Genes from Cuphea paucipetala, Cupheaignea, Cuphea painteri, and Cuphea hookeriana

Lysophosphatidic acyltransferase (LPAAT) genes from plant seeds werecloned and expressed in the transgenic strain, S6511, derived from UTEX1435 (P. moriformis). Expression of the heterologous LPAATs increasesC8:0 and C10:0 fatty acid levels and dramatically increasesincorporation of C8:0 and C10:0 fatty acids at the sn-2 position oftriacylglycerols (TAGs) in transgenic strains.

TAGs are synthesized from various chain length acyl-CoAs andglycerol-3-phosphate by consecutive action of three ER-resident enzymesof the Kennedy pathway-glycerol phosphate acyltransferase (GPAT), LPAAT,and diacylglycerol acyltransferase (DGAT). Substrate specificities ofthese acyltransferases are known to determine the fatty acid compositionof the resulting TAGs. LPAAT acylates the sn-2 hydroxyl group oflysophosphatidic acid (LPA) to form phosphatidic acid (PA), a precursorto TAG. In co-owned applications WO2013/158938, WO2015/051139, andPCT/US2016/026265 we demonstrated expression of LPAAT from Cocosnucifera (CnLPAAT, accession no. AAC49119; Knutzon et al., 1995).

Strain S6511 expresses the acyl-ACP thioesterase (FATB2) gene fromCuphea hookeriana (ChFATB2), leading to C8:0 and C10:0 fatty acidaccumulation of ca. 14% and 28%, respectively. Strain S6511 is a strainmade according to the methods disclosed in co-owned WO2010/063031 andWO2010/063032, herein incorporated by reference. Briefly, S6511 is astrain that express sucrose invertase and a C. hookeriana FATB2. Theconstruct pSZ3101:6S::CrTUB2-ScSUC2-CvNR_a:PmAMT03-CpSAD1tp_trimmed:ChFATB2-CvNR_d::6S wasengineered into S3150, a strain classically mutagenized to increaselipid yield. We identified novel C8:0- and C10:0-specific LPAATs fromseeds exhibiting high levels of C8:0 and C10:0 fatty acids. After weidentified and cloned LPAATs we expressed the LPAAT genes in S6511.

Method for Identification of LPAATs

Seeds were obtained from species exhibiting elevated levels of midchainand other specialized fatty acids (Table 4).

TABLE 4 Fatty acid profiles of mature seeds. C18:1 C22: C8: C10: C12:C14: C16: C18: C18: (petro- C18: C20: C20: C22: C22: C22: 2n9, C22: 0 00 0 0 0 1 selinate) 2 0 1 0 1n17 1n9 17 2n6 S01_Cc Cinnamomum 0.4 54.739.0 1.6 0.7 0.1 2.9 0.6 0.0 camphora S02_Uc Umbellularia 0.9 28.8 63.02.3 0.4 0.1 3.4 0.6 0.0 californica S03_Ld Limnanthes 0.0 0.0 0.0 0.40.7 0.4 2.7 1.5 1.5 59.9 0.3 2.8 17.4 9.3 0.5 douglasii S04_Chs Cuphea0.2 6.5 83.7 5.1 1.1 0.1 0.0 1.7 0.1 hyssopifolia S05_Ccr Cuphea 1.6 8.159.2 15.2 3.9 0.6 0.0 5.4 0.2 carthagenensis S06_Cpr Cuphea 2.0 11.561.3 10.8 2.7 0.5 0.0 5.2 0.1 parsonsia S07_Cg Cuphia 7.1 85.1 1.7 0.31.0 0.2 0.0 2.1 0.1 glossostoma S08_Cht Cuphea 3.5 44.3 40.0 4.3 1.2 0.32.2 3.6 0.1 heterophylla S11_Dc Daucus 0.0 0.0 0.0 0.1 5.9 0.8 11.5 65.913.0 0.5 0.3 0.3 carrota S14_Cw Cuphea 0.5 20.2 62.5 5.8 2.2 0.3 2.7 4.7wrightii S15_Bj Brassica 0.0 0.0 0.0 0.1 3.2 0.7 12.1 19.2 0.5 6.3 0.838.9 1.3 juncea S16_Br Brassica 0.0 0.0 0.0 0.1 2.8 1.0 16.0 16.8 0.78.3 1.0 40.4 0.8 rapa nipposinica S17_Ca Cuphea 90.8 2.7 0.0 0.1 1.2 0.11.8 2.8 avigera var. pulcherrima S18_Ch Cuphea 64.7 29.7 0.1 0.2 1.3 0.11.9 2.0 hookeriana S19_Cpal Cuphea 28.9 0.8 1.3 55.1 6.2 0.2 3.0 3.4palustris S20_Cpai Cuphea 67.0 20.8 0.1 0.2 2.6 0.3 3.1 4.5 painteriS21_Cpau Cuphea 1.5 91.0 1.2 0.7 1.5 0.2 1.1 2.1 paucipetala S22_ChookCuphea 62.8 31.9 0.2 0.2 1.0 0.1 2.1 1.2 hookeriana S23_Cglut Cuphea 5.229.9 46.4 3.9 1.9 0.4 0.0 8.1 glutinosa S24_Caequ Cuphea 27.1 0.0 1.457.4 6.0 0.2 3.2 3.8 aequipetala S25_Ccalc Cuphea 8.0 20.4 46.8 7.6 3.20.6 3.7 8.5 calcarata S26_Chook Cuphea 70.4 23.1 0.1 0.2 1.5 0.2 2.5 1.8hookeriana S27_Cproc Cuphea 0.9 86.3 0.0 1.6 2.2 0.4 3.2 3.3 procumbensS28_Cignea Cuphea 3.1 84.9 0.7 0.3 2.6 0.2 2.9 4.4 ignea S35_CcrasCuphea 1.3 87.7 1.3 0.4 2.0 0.5 3.3 2.7 crassiflora S36_Ckoe Cuphea 0.087.4 1.4 0.8 2.2 0.4 2.3 4.5 koehneana S37_Clept Cuphea 1.3 86.1 1.3 0.42.2 0.5 3.1 4.1 leptopoda S40_Clop Cuphea 0.5 82.3 2.4 1.6 3.0 0.6 3.94.9 lophostoma S41_Sal Sassafras 4.3 65.2 22.8 0.9 0.8 5.1 0.0 0.6albidum db The percentage of each fatty acid making up the seed oil isshown; abundant and unusual fatty acid species are indicated in bold.

Briefly, RNA was extracted from dried plant seeds and submitted forpaired-end sequencing using the Illumina Hiseq 2000 platform. RNAsequence reads were assembled into corresponding seed transcriptomesusing the Trinity software package. LPAAT-containing cDNA contigs wereidentified by mining transcriptomes for sequences with homology to aknown LPAAT that was previously identified in-house, CuPSR23 LPAAT2-1(see WO2013/158938), using BLAST. For some sequences, a high-confidence,full-length transcript was assembled using Trinity. The resulting aminoacid sequences of all new LPAATs were subjected to phylogenetic analysesusing previously known, full-length LPAAT sequences (available via NCBI)as well as sequences of previously known LPAATs whose sequences werederived at Solazyme. The analysis showed that the amino acid sequencesof the newly discovered LPPAATs were not similar to previously knownLPAATs. Table 5 shows the clade analysis in which the novel LPAATs wereclustered according to a neighbor joining algorithm. These were found toform 4 clades as listed in Table 5.

TABLE 5 Clade Analysis of LPAATs Percent amino acid Amino Acid identityClade SEQ ID Nos. to members No. in Clade Full Genus Species Function ofclade 1 S15 BjLPAAT1d Brassica juncea 96.3 S15 BjLPAAT1c Brassica junceaS15 BjLPAAT1a Brassica juncea S15 BjLPAAT1b Brassica juncea 2CuPSR23LPAAT2-1 Cuphea PSR23 Prefer C8/ 93.9 S40 ClopLPAAT1 Cuphealophostoma C10 sn-2 S21 CpauLPAAT1 Cuphea paucipetala S37 CleptLPAAT1Cuphea leptopoda S27 CprocLPAAT1b Cuphea procumbens S27 CprocLPAAT1Cuphea procumbens S04 ChsLPAAT2 Cuphea hyssopifolia S28 CigneaLPAAT1Cuphea ignea S05 CcrLPAAT2a Cuphea carthagenensis S06 CprLPAAT1 Cupheaparsonsia S05 CcrLPAAT2b Cuphea carthagenensis S17 CaLPAAT3 Cupheaavigera var. pulcherrima S26 ChookLPAAT1 Cuphea hookeriana S20CpaiLPAAT1 Cuphea painteri S04 ChsLPAAT1 Cuphea hyssopifolia S25 Ccalc1aCuphea calcarata S25 Ccalc1b Cuphea calcarata S14 CwLPAAT1 Cupheawrightii S08 ChtLPAAT1a Cuphea heterophylla S08 ChtLPAAT1b Cupheaheterophylla S36 CkoeLPAAT2 Cuphea koehneana S02 UcLPAAT1b Umbellulariacalifornica S02 UcLPAAT1a Umbellularia californica S01 CcLPAAT1aCinnamomum camphora S01 CcLPAAT1b Cinnamomum camphora S41 SaILPAAT1Sassafras albidum db 3 S14 CwLPAAT2a Cuphea wrightii C18:2 86.5 S14CwLPAAT2b Cuphea wrightii S25 CcalcLPAAT2 Cuphea calcarata S19CpaILPAAT1 Cuphea palustris S22 ChookLPAAT3b Cuphea hookeriana S17CaLPAAT1 Cuphea avigera var. pulcherrima S22 ChookLPAAT3a Cupheahookeriana CuPSR23LPAAT3-1 Cuphea PSR23 S27 CprocLPAAT2b Cupheaprocumbens S27 CprocLPAAT2a Cuphea procumbens S18 ChLPAAT2a Cupheahookeriana S24 CaequLPAAT1d Cuphea aequipetala S24 CaequLPAAT1b Cupheaaequipetala S24 CaequLPAAT1a Cuphea aequipetala S24 CaequLPAAT1c Cupheaaequipetala S23 CglutLPAAT1a Cuphea glutinosa S23 CglutLPAAT1b Cupheaglutinosa S26 ChookLPAAT2b Cuphea hookeriana S07 CgLPAAT1c Cuphiaglossostoma S07 CgLPAAT1b Cuphia glossostoma S07 CgLPAAT1a Cuphiaglossostoma S28 CigneaLPAAT2 Cuphea ignea S36 CkoeLPAAT1 Cupheakoehneana S35 CcrasLPAAT1a Cuphea crassiflora S35 CcrasLPAAT1c Cupheacrassiflora S35 CcrasLPAAT1b Cuphea crassiflora S35 CcrasLPAAT1d Cupheacrassiflora 4 Gh LPAAT2B Garcinia hombroriana Reduced 78.5 Gi LPAAT2B-1Garcinia indica trisaturates, Gh LPAAT2A Garcinia hombroriana increaseGi LPAAT2A Garcinia indica unsaturates Gh LPAAT2C Garcinia hombrorianaat Sn-2 Gi LPAAT2C-2 Garcinia indica position S03 LdLPAAT1 Limnanthesdouglasii S11 DcLPAAT1 Daucus carrota (carrot) S11 DcLPAAT2 Daucuscarrota (carrot) S11 DcLPAAT2 Daucus carrota (truncated) (carrot)Functionality of LPAATs in P. moriformis

To increase the levels of C8:0 and C10:0 fatty acids in strain S6511, aswell as to test the functionality of the newly identified LPAATs, weidentified midchain-specific LPAATs from the transcriptomes of speciesexhibiting high levels of C8:0 and C10:0 fatty acids in their oil seedsand introduced the genes into S6511. LPAATs that co-clustered withCuPSR23 LPAAT2-1, specifically CpauLPAAT1, CigneaLPAAT1, ChookLPAAT1,and CpaiLPAAT1, were selected for synthesis and testing. CpauLPAAT1,CigneaLPAAT1, ChookLPAAT1, and CpaiLPAAT1 were synthesized in acodon-optimized form to reflect UTEX 1435 codon usage. Transgenicstrains were generated via transformation of the strain S6511 with aconstruct encoding one of the four LPAAT genes. The construct pSZ3840encoding CpauLPAAT1 is shown as an example, but identical methods wereused to generate each of the remaining three constructs. ConstructpSZ3840 can be written aspLOOP::PmHXT1-ScarMEL1-CvNR:PmAMT3-CpauLPAAT1-CvNR::pLOOP. The sequenceof the transforming DNA is provided in FIG. 2 (pSZ3840). The relevantrestriction sites in the construct from 5′-3′, BspQI, KpnI, SpeI, XhoI,EcoRI, SpeI, XhoI, SacI, BspQI, respectively, are indicated inlowercase, bold, and underlined. BspQI sites delimit the 5′ and 3′ endsof the transforming DNA. Bold lowercase sequences at the 5′ and 3′ endof the construct represent genomic DNA from UTEX 1435 that targetintegration to the pLOOP locus via homologous recombination. Proceedingin the 5′ to 3′ direction, the selection cassette has the P. moriformisHXT1 promoter driving expression of the Saccharomyces carlsbergensisMEL1 (conferring the ability to grow on melibiose) and the Chlorellavulgaris Nitrate reductase (NR) gene 3′ UTR. The promoter is indicatedby lowercase, boxed text. The initiator ATG and terminator TGA forScarMEL1 are indicated in bold, uppercase italics, while the codingregion is indicated with lowercase italics. The 3′ UTR is indicated bylowercase underlined text. The second cassette containing the codonoptimized CpauLPAAT1 gene from Cuphea paucipetala is driven by the P.moriformis AMT3 promoter and has the Chlorella vulgaris Nitratereductase (NR) gene 3′ UTR. In this cassette, the AMT3 promoter isindicated by lowercase, boxed text. The initiator ATG and terminator TGAfor the CpauLPAAT1 gene are indicated in bold, uppercase italics, whilethe coding region is indicated by lowercase italics. The 3′ UTR isindicated by lowercase underlined text. The final construct wassequenced to ensure correct reading frame and targeting sequences.

SEQ ID NO: 19 pSZ3840/D2554 transforming construct (CpauLPAAT1)

aatacaatattcagtatgtcgcgggcggcgacggcggggagctgatgtcgcgctgggtattgcttaatcgccagcttcgcccccgtcttggcgcgaggcgtgaacaagccgaccgatgtgcacgagcaaatcctgacactagaagggctgactcgcccggcacggctgaattacacaggcttgcaaaaataccagaatttgcacgcaccgtattcgcggtattttgttggacagtgaatagcgatgcggcaatggcttgtggcgttagaaggtgcgacgaaggtggtgccaccactgtgccagccagtcctggcggctcccagggccccgatcaagagccaggacatccaaactacccacagcatcaacgccccggcctatactcgaaccccacttgcactctgcaatggtatgggaaccacgggg

gcctgacgccccagatgggctgggacaactggaacacgttcgcctgcgacgtctccgagcagctgctgctggacacggccgaccgcatctccgacctgggcctgaaggacatgggctacaagtacatcatcctggacgactgctggtcctccggccgcgactccgacggcttcctggtcgccgacgagcagaagttccccaacggcatgggccacgtcgccgaccacctgcacaacaactccttcctgttcggcatgtactcctccgcgggcgagtacacgtgcgccggctaccccggctccctgggccgcgaggaggaggacgcccagttcttcgcgaacaaccgcgtggactacctgaagtacgacaactgctacaacaagggccagttcggcacgcccgagatctcctaccaccgctacaaggccatgtccgacgccctgaacaagacgggccgccccatcttctactccctgtgcaactggggccaggacctgaccttctactggggctccggcatcgcgaactcctggcgcatgtccggcgacgtcacggcggagttcacgcgccccgactcccgctgcccctgcgacggcgacgagtacgactgcaagtacgccggcttccactgctccatcatgaacatcctgaacaaggccgcccccatgggccagaacgcgggcgtcggcggctggaacgacctggacaacctggaggtcggcgtcggcaacctgacggacgacgaggagaaggcgcacttctccatgtgggccatggtgaagtcccccctgatcatcggcgcgaacgtgaacaacctgaaggcctcctcctactccatctactcccaggcgtccgtcatcgccatcaaccaggactccaacggcatccccgccacgcgcgtctggcgctactacgtgtccgacacggacgagtacggccagggcgagatccagatgtggtccggccccctggacaacggcgaccaggtcgtggcgctgctgaacggcggctccgtgtcccgccccatgaacacgaccctggaggagatcttcttcgactccaacctgggctccaagaagctgacctccacctgggacatctacgacctgtgggcgaaccgcgtcgacaactccacggcgtccgccatcctgggccgcaacaagaccgccaccggcatcctgtacaacgccaccgagcagtcctacaaggacggcctgtccaagaacgacacccgcctgttcggccagaagatcggctccctgtcccccaacgcgatcctgaacacgaccgtccccgcccacggcatcgcgttctaccgcctgcgcccc

acacttgctgccttgacctgtgaatatccctgccgcttttatcaaacagcctcagtgtgtttgatcttgtgtgtacgcgcttttgcgagttgctagctgcttgtgctatttgcgaataccacccccagcatccccttccctcgtttcatatcgcttgcatcccaaccgcaacttatctacgctgtcctgctatccctcagcgctgctcctgctcctgctcactgcccctcgcacagccttggtttgggctccgcctgtattctcctggtact

atcaacctgttccaggccctgtgcttcgtgctggtgtggcccctgtccaagaacgcctaccgccgcatcaaccgcgtgttcgccgagctgctgctgtccgagctgctgtgcctgttcgactggtgggccggcgccaagctgaagctgttcaccgaccccgagaccttccgcctgatgggcaaggagcacgccctggtgatcatcaaccacatgaccgagctggactggatgctgggctgggtgatgggccagcacctgggctgcctgggctccatcctgtccgtggccaagaagtccaccaagttcctgcccgtgctgggctggtccatgtggttctccgagtacctgtacatcgagcgctcctgggccaaggaccgcaccaccctgaagtcccacatcgagcgcctgaccgactaccccctgcccttctggatggtgatcttcgtggagggcacccgcttcacccgcaccaagctgctggccgcccagcagtacgccgcctcctccggcctgcccgtgccccgcaacgtgctgatcccccgcaccaagggcttcgtgtcctgcgtgtcccacatgcgctccttcgtgcccgccgtgtacgacgtgaccgtggccttccccaagacctcccccccccccaccctgctgaacctgttcgagggccagtccatcgtgctgcacgtgcacatcaagcgccacgccatgaaggacctgcccgagtccgacgacgccgtggcccagtggtgccgcgacaagttcgtggagaaggacgccctgctggacaagcacaacgccgaggacaccttctccggccaggaggtgcaccgcaccggctcccgccccatcaagtccctgctggtggtgatctcctgggtggtggtgatcaccttcggcgccctgaagttcctgcagtggtcctcctggaagggcaaggccttctccgtgatcggcctgggcatcgtgaccctgctgatgcacatgctgatcctgtcctcccaggccgagcgctc

gacctgtgaatatccctgccgcttttatcaaacagcctcagtgtgtttgatcttgtgtgtacgcgcttttgcgagttgctagctgcttgtgctatttgcgaataccacccccagcatccccttccctcgtttcatatcgcttgcatcccaaccgcaacttatctacgctgtcctgctatccctcagcgctgctcctgctcctgctcactgcccctcgcacagccttggtttgggctccgcctgtattctcctggtactgcaacctgtaaac

gtacgacgttgggcacgcccatgaaagtttgtataccgagcttgttgagcgaactgcaagcgcggctcaaggatacttgaactcctggattgatatcggtccaataatggatggaaaatccgaacctcgtgcaagaactgagcaaacctcgttacatggatgcacagtcgccagtccaatgaacattgaagtgagcgaactgttcgcttcggtggcagtactactcaaagaatgagctgctgttaaaaatgcactctcgttctctcaagtgagtggcagatgagtgctcacgccttgcacttcgctgcccgtgtcatgccctgcgccccaaaatttgaaaaaagggatgagattattgggcaatggacgacgtcgtcgctccgggagtcaggaccggcggaaaataagaggcaacacactccgcttctt

The sequence for all of the other LPAAT constructs are identical to thatof pSZ3840 with the exception of the encoded LPAAT. The LPAAT sequencealone with flanking SpeI and XhoI restriction sites is provided for theremaining LPAAT constructs are shown below. The amino acid sequence ofthe LPAAT proteins is provided below.

pSZ3841/D2555 (CpaiLPAAT1) SEQ ID NO: 20 actagt

gccatcccctccgccgccgtggtgttcctgttcggcctgctgttcttcacctccggcctgatcatcaacctgttccaggccttctgcttcgtgctgatctcccccctgtccaagaacgcctaccgccgcatcaaccgcgtgacgccgagctgctgcccctggagacctgtggctgttccactggtgcgccggcgccaagctgaagctgttcaccgaccccgagaccttccgcctgatgggcaaggagcacgccctggtgatcatcaaccacaagatcgagctggactggatggtgggctgggtgctgggccagcacctgggctgcctgggctccatcctgtccgtggccaagaagtccaccaagttcctgcccgtgttcggctggtccctgtggttctccggctacctgttcctggagcgctcctgggccaaggacaagatcaccctgaagtcccacatcgagtccctgaaggactaccccctgcccttctggctgatcatcttcgtggagggcacccgcttcacccgcaccaagctgctggccgcccagcagtacgccgcctcctccggcctgcccgtgccccgcaacgtgctgatcccccacaccaagggcttcgtgtcctccgtgtcccacatgcgctccttcgtgcccgccatctacgacgtgaccgtggccttccccaagacctcccccccccccaccatgctgaagctgacgagggccagtccgtggagctgcacgtgcacatcaagcgccacgccatgaaggacctgcccgagtccgacgacgccgtggcccagtggtgccgcgacaagttcgtggagaaggacgccctgctggacaagcacaactccgaggacaccttctccggccaggaggtgcaccacgtgggccgccccatcaaggccctgctggtggtgatctcctgggtggtggtgatcatcttcggcgccctgaagttcctgctgtggtcctccctgctgtcctcctggaagggcaaggccactccgtgatcggcctgggcatcgtggccggcatcgtgaccctgctgatgcacatcctgatcctgtcctcccaggccgagggctccaaccccgtgaaggccgcccccgccaagctgaagaccgagctgtcctcctccaagaaggtgaccaacaaggagaac

ctcgag pSZ3842/D2556 (CigneaLPAAT1) SEQ ID NO: 21 actagt

gccatcgccgccgccgccgtgatcttcctgttcggcctgctgttcttcgcctccggcatcatcatcaacctgttccaggccctgtgcttcgtgctgatctggcccctgtccaagaacgtgtaccgccgcatcaaccgcgtgacgccgagctgctgctgatggacctgctgtgcctgttccactggtgggccggcgccaagatcaagctgacaccgaccccgagaccttccgcctgatgggcatggagcacgccctggtgatcatgaaccacaagaccgacctggactggatggtgggctggatcctgggccagcacctgggctgcctgggctccatcctgtccatcgccaagaagtccaccaagttcatccccgtgctgggctggtccgtgtggactccgagtacctgttcctggagcgctcctgggccaaggacaagtccaccctgaagtcccacatggagaagctgaaggactaccccctgcccttctggctggtgatcttcgtggagggcacccgcttcacccgcaccaagctgctggccgcccagcagtacgccgcctcctccggcctgcccgtgccccgcaacgtgctgatcccccacaccaagggcttcgtgtcctgcgtgtccaacatgcgctccacgtgcccgccgtgtacgacgtgaccgtggccttccccaagtcctcccccccccccaccatgctgaagctgttcgagggccagtccatcgtgctgcacgtgcacatcaagcgccacgccctgaaggacctgcccgagtccgacgacgccgtggcccagtggtgccgcgacaagttcgtggagaaggacgccctgctggacaagcacaacgccgaggacaccttctccggccaggaggtgcaccacatcggccgccccatcaagtccctgctggtggtgatcgcctgggtggtggtgatcatcttcggcgccctgaagttcctgcagtggtcctccctgctgtccacctggaagggcaaggccttctccgtgatcggcctgggcatcgccaccctgctgatgcacatgctgatcctgtcctcccaggccgagcgctccaaccccgccaaggtggccaag

ctcgag pSZ3844/D2557 (ChookLPAAT1) SEQ ID NO: 22 actagt

gccatcccctccgccgccgtggtgttcctgttcggcctgctgttcttcacctccggcctgatcatcaacctgttccaggccttctgcttcgtgctgatctcccccctgtccaagaacgcctaccgccgcatcaaccgcgtgacgccgagctgctgcccctggagacctgtggctgttccactggtgcgccggcgccaagctgaagctgttcaccgaccccgagaccttccgcctgatgggcaaggagcacgccctggtgatcatcaaccacaagatcgagctggactggatggtgggctgggtgctgggccagcacctgggctgcctgggctccatcctgtccgtggccaagaagtccaccaagttcctgcccgtgttcggctggtccctgtggttctccgagtacctgttcctggagcgctcctgggccaaggacaagatcaccctgaagtcccacatcgagtccctgaaggactaccccctgcccttctggctgatcatcttcgtggagggcacccgcttcacccgcaccaagctgctggccgcccagcagtacgccgcctcctccggcctgcccgtgccccgcaacgtgctgatcccccacaccaagggcttcgtgtcctccgtgtcccacatgcgctccttcgtgcccgccatctacgacgtgaccgtggccttccccaagacctcccccccccccaccatgctgaagctgacgagggccagtccgtggagctgcacgtgcacatcaagcgccacgccatgaaggacctgcccgagtccgacgacgccgtggcccagtggtgccgcgacaagttcgtggagaaggacgccctgctggacaagcacaactccgaggacaccttctccggccaggaggtgcaccacgtgggccgccccatcaaggccctgctggtggtgatctcctgggtggtggtgatcatcttcggcgccctgaagttcctgctgtggtcctccctgctgtcctcctggaagggcaaggccactccgtgatcggcctgggcatcgtggccggcatcgtgaccctgctgatgcacatcctgatcctgtcctcccaggccgagggctccaaccccgtgaaggccgcccccgccaagctgaagaccgagctgtcctcctccaagaaggtgaccaacaaggagaac

ctcgag

To determine the impact of the CpauLPAAT1, CigneaLPAAT1, ChookLPAAT1,and CpaiLPAAT1 genes on mid-chain fatty acid accumulation, the aboveconstructs containing the codon optimized CpauLPAAT1, CigneaLPAAT1,ChookLPAAT1, and CpaiLPAAT1 genes were transformed into strain S6511.Primary transformants were clonally purified and grown under standardlipid production conditions at pH7.0 (all the strains require growth atpH 7.0 to allow for maximal expression of the LPAAT gene driven by thepH-regulated AMT3 promoter). The resulting profiles from a set ofrepresentative clones arising from these transformations are shown inTable 6.

TABLE 6 Transformants of pSZ3840 (CpauLPAAT1), pSZ3841 (CpaiLPAAT1),pSZ3842 (CigneaLPAAT1), and pSZ3844 (ChookLPAAT1). The fatty acidprofiles for transgenic strains expressing LPAATs derived from C.paucipetala, C. painteri, C. ignea, and C. hookeriana. Sample ID C8:0C10:0 C12:0 C14:0 C16:0 C18:0 C18:1 C18:2 C18:3 a Parent S6511a 14.427.7 0.6 1.3 8.8 1.6 38.2 5.4 0.4 S6511b 14.5 27.7 0.6 1.3 8.6 1.6 38.45.3 0.4 pSZ3840 CpauLPAAT1 S6511; T792; D2554-20 16.6 29.9 0.7 1.3 8.01.0 35.2 5.2 0.5 S6511; T792; D2554-17 14.6 28.7 0.6 1.3 8.4 1.7 37.15.7 0.5 S6511; T792; D2554-41 15.2 28.5 0.7 1.3 8.3 1.4 37.5 5.2 0.4S6511; T792; D2554-35 14.7 28.4 0.6 1.3 8.6 1.6 37.3 5.6 0.5 S6511;T792; D2554-27 15.2 27.6 0.7 1.3 9.5 1.5 37.1 5.1 0.4 pSZ3841 CpaiLPAAT1S6511; T792; D2555-34 17.3 29.5 0.7 1.3 7.8 1.2 35.1 5.1 0.4 S6511;T792; D2555-43 17.5 29.1 0.7 1.3 8.0 0.9 35.4 5.0 0.5 S6511; T792;D2555-10 15.7 28.3 0.7 1.3 8.6 1.6 36.2 5.7 0.5 S6511; T792; D2555-2216.0 27.9 0.7 1.3 8.4 0.9 37.8 5.0 0.4 S6511; T792; D2555-44 15.3 27.50.6 1.3 8.1 1.8 38.2 5.4 0.4 pSZ3842 CigneaLPAAT1 S6511; T792; D2556-3816.2 29.2 0.7 1.3 8.1 1.3 36.1 5.2 0.5 S6511; T792; D2556-22 14.3 28.50.7 1.3 8.5 1.6 37.6 5.7 0.5 S6511; T792; D2556-44 13.6 28.4 0.7 1.4 9.01.5 36.3 6.7 0.7 S6511; T792; D2556-14 14.1 28.0 0.6 1.3 8.6 1.7 38.05.6 0.5 S6511; T792; D2556-36 14.3 28.0 0.6 1.3 8.6 1.7 37.9 5.7 0.5pSZ3844 ChookLPAAT1 S6511; T792; D2557-47 15.8 29.3 0.7 1.3 8.2 1.2 36.55.0 0.5 S6511; T792; D2557-24 16.8 28.8 0.7 1.3 8.1 1.2 35.8 5.4 0.5S6511; T792; D2557-30 15.2 28.3 0.7 1.3 8.5 1.6 36.8 5.7 0.5 S6511;T792; D2557-39 14.7 28.2 0.7 1.3 8.7 1.5 37.3 5.7 0.5 S6511; T792;D2557-26 15.3 27.7 0.7 1.4 8.7 0.9 37.7 5.4 0.5

The transformants in Table 6 display a marked increase in the productionof C8:0 and C10:0 fatty acids upon expression of the heterologousLPAATs. To determine if expression of the heterologous LPAAT genesaffected the regiospecificity of fatty acids at the sn-2 position, weanalyzed TAGs from representative D2554 (CpauLPAAT1), D2555(CpaiLPAAT1), D2556 (CigneaLPAAT1), and D2557 (ChookLPAAT1) strainsutilizing the porcine pancreatic lipase method. Cells were grown underconditions to maximize midchain fatty acid levels and to generatesufficient biomass for TAG analysis. TAG and sn-2 profiles are shown inTable 7.

Table 7:

Inclusion of C8:0 and C10:0 fatty acids at the sn-2 position of TAGs.Selected transformants were subjected to porcine pancreatic lipasedetermination of fatty acid inclusion at the sn-2 position. The generalfatty acid distribution in triacylglycerols (TAG) is shown to indicatefatty acid abundance for each transformant. In addition, thesn-2-specific distribution is shown. Numbers highlighted in bold anditalic reflect significantly increased inclusion of the noted fatty acidcompared to the parent S6511.

TABLE 7 S6511; T792; S6511; T792; S6511; T792; S6511; T792; D2554-20D2555-34 D2556-38 D2557-24 Strain: S6511 (CpauLPAAT1) (CpaiLPAAT1)(CigneaLPAAT1) (ChookLPAAT1) Analysis TAG sn-2 TAG sn-2 TAG sn-2 TAGsn-2 TAG sn-2 Fatty Acid C8:0 14.4 8.5 16.6 12.8 17.3 22.3 16.2 10.016.8 29.1 (area %) C10:0 27.7 26.4 29.9 39.0 29.5 22.2 29.2 36.2 28.819.4 C12:0 0.6 0.4 0.7 0.3 0.7 0.4 0.7 0.4 0.7 0.3 C14:0 1.3 1.0 1.3 1.01.3 0.9 1.3 1.2 1.3 0.9 C16:0 8.8 0.9 8.0 1.1 7.8 1.1 8.1 1.2 8.1 0.9C18:0 1.6 0.2 1.0 0.4 1.2 0.5 1.3 0.5 1.2 0.3 C18:1 38.2 52.5 35.2 37.835.1 43.6 36.1 42.2 35.8 40.7 C18:2 5.4 8.9 5.2 6.2 5.1 7.9 5.2 7.0 5.47.1 C18:3 α 0.4 0.8 0.5 0.7 0.4 0.9 0.5 0.8 0.5 0.7 C8 + C10 42.2 34.946.4 51.8 46.8 44.5 45.5 46.1 45.6 48.5 sum

As disclosed in Table 7, the CpauLPAAT1 and CigneaLPAAT1 genes showremarkable specificity towards C10:0 fatty acids. D2554-20 exhibits39.0% of C10:0 in the sn-2 position versus just 26.4% in the S6511 basestrain without the heterologous LPAAT, demonstrating a 1.5 fold increasein C10:0 inclusion at the sn-2 position. D2556-38 exhibits 36.2% ofC10:0 in the sn-2 position versus 26.4% in the S6511 base strain,demonstrating a 1.4 fold increase in C10:0 inclusion at the sn-2position. Although there is a small increase in C8:0 levels in theD2554-20 and D2555-34 strains, the vast majority of sn-2 targeting isC10:0-specific. Similarly, CpaiLPAAT1 and ChookLPAAT1 show remarkablespecificity towards C8:0 fatty acids. D2555-34 exhibits 22.3% C8:0 inthe sn-2 position versus just 8.5% in the S6511 base strain without theheterologous LPAAT, demonstrating a 2.6 fold increase in C8:0 inclusionat the sn-2 position. D2557-24 exhibits 29.1% C8:0 in the sn-2 positionversus 8.5%, demonstrating a 3.4 fold increase in C8:0 inclusion at thesn-2 position. We teach that CpauLPAAT1 and CigneaLPAAT1 areC10:0-specific LPAATs and that CpaiLPAAT1 and ChookLPAAT1 areC8:0-specific LPAATs. Knutzon D S, Lardizabal K D, Nelsen J S, BleibaumJ L, Davies H M, Metz J G (1995) Cloning of a coconut endosperm cDNAencoding a 1-acyl-sn-glycerol-3-phosphate acyltransferase that acceptsmedium-chain-length substrates. Plant Physiol 109:999-1006

Amino Acid Sequences for Novel LPAAT Genes

CpauLPAAT1 SEQ ID NO: 23MAIPAAAVIFLFGLLFFTSGLIINLFQALCFVLVWPLSKNAYRRINRVFAELLLSELLCLFDWWAGAKLKLFTDPETFRLMGKEHALVIINHMTELDWMLGWVMGQHLGCLGSILSVAKKSTKFLPVLGWSMWFSEYLYIERSWAKDRTTLKSHIERLTDYPLPFWMVIFVEGTRFTRTKLLAAQQYAASSGLPVPRNVLIPRTKGFVSCVSHMRSFVPAVYDVTVAFPKTSPPPTLLNLFEGQSIVLHVHIKRHAMKDLPESDDAVAQWCRDKFVEKDALLDKHNAEDTFSGQEVHRTGSRPIKSLLVVISWVVVITFGALKFLQWSSWKGKAFSVIGLGIVTLLMHMLILSSQAERSSNPAKVAQAKLKTELSISKKATDKEN CprocLPAAT1SEQ ID NO: 24 MAIPAAAVIFLFGLIFFASGLIINLFQALCFVLIWPISKNAYRRINRVFAELLLSELLCLFDWWAGAKLKLFTDPETFRLMGKEHALVIINHMTELDWMVGWVMGQHFGCLGSILSVAKKSTKFLPVLGWSMWFTEYLYIERSWNKDKSTLKSHIERLKDYPLPFWLVIFAEGTRFTQTKLLAAQQYAASSGLPVPRNVLIPRTKGFVSCVSHMRSFVPAVYDLTVAFPKTSPPPTLLNLFEGQSVVLHVHIKRHAMKDLPESDDEVAQWCRDKFVEKDALLDKHNAEDTFSGQELQHTGRRPIKSLLVVISWVVVIAFGALKFLQWSSWKGKAFSVIGLGIVTLLMHMLILSSQAERSKPAKVAQAKLKTELSISKTVTDKEN CprocLPAAT1b SEQ ID NO: 25MAIPAAAVIFLFGLIFFASGLIINLFQALCFVLIWPISKNAYRRINRVFAELLLSELLCLFDWWAGAKLKLFTDPETFRLMGKEHALVIINHMTELDWMVGWVMGQHFGCLGSILSVAKKSTKFLPVLGWSMWFTEYLYIERSWNKDKSTLKSHIERLKDYPLPFWLVIFAEGTRFTQTKLLAAQQYAASSGLPVPRNVLIPRTKGFVSCVSHMRSFVPAVYDLTVAFPKTSPPPTLLNLFEGQSVVLHVHIKRHAMKDLPESDDEVAQWCRDKFVEK CprocLPAAT2a SEQ ID NO: 26IVNLVQAVCFVLVRPLSKNTYRRINRVVAELLWLELVWLIDWWAGVKIKVFTDHETFHLMGKEHALVICNHKSDIDWLVGWVLAQRSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYLFLERNWAKDESTLKSGLNRLKDYPLPFWLALFVEGTRFTRAKLLAAQQYAASSGLPVPRNVLIPRTKGFVSSVSHMRSFVPAIYDVTVAIPKTSPPPTLIRMFKGQSSVLHVHLKRHVMKDLPESDDAVAQWCRDIFVEKDALLDKHNADDTFSGQELQDTGRPIKSLLVVISWAVLEVFGAVKFLQWSSLLSSWKGLAFSGIGLGIITLLMHILILFSQSERSTPAKVAPAKAKIEGESSKTEMEKEK CprocLPAAT2b SEQ ID NO: 27 IVNLVQAVCFVLVRPLSKNTYRRINRVVAELLWLELVWLIDWWAGVKIKVFTDHETFHLMGKEHALVICNHKSDIDWLVGWVLAQRSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYLFLERNWAKDESTLKSGLNRLKDYPLPFWLALFVEGTRFTRAKLLAAQQYAASSGLPVPRNVLIPRTKGFVSSVSHMRSFVPAIYDVTVAIPKTSPPPTLIRMFKGQSSVLHVHLKRHVMKDLPESDDAVAQWCRDIFVEKDALLDKHNADDTFSGQELQDTGRPIKSLLV CpaiLPAAT1 SEQ ID NO: 28 MAIPSAAVVFLFGLLFFTSGLIINLFQAFCFVLISPLSKNAYRRINRVFAELLPLEFLWLFHWCAGAKLKLFTDPETFRLMGKEHALVIINHKIELDWMVGWVLGQHLGCLGSILSVAKKSTKFLPVFGWSLWFSGYLFLERSWAKDKITLKSHIESLKDYPLPFWLIIFVEGTRFTRTKLLAAQQYAASSGLPVPRNVLIPHTKGFVSSVSHMRSFVPAIYDVTVAFPKTSPPPTMLKLFEGQSVELHVHIKRHAMKDLPESDDAVAQWCRDKFVEKDALLDKHNSEDTFSGQEVHHVGRPIKALLVVISWVVVIIFGALKFLLWSSLLSSWKGKAFSVIGLGIVAGIVTLLMHILILSSQAEGSNPVKAAPAKLKTELSSSKKVTNKEN ChookLPAAT1SEQ ID NO: 29  MAIPSAAVVFLFGLLFFTSGLIINLFQAFCFVLISPLSKNAYRRINRVFAELLPLEFLWLFHWCAGAKLKLFTDPETFRLMGKEHALVIINHKIELDWMVGWVLGQHLGCLGSILSVAKKSTKFLPVFGWSLWFSEYLFLERSWAKDKITLKSHIESLKDYPLPFWLIIFVEGTRFTRTKLLAAQQYAASSGLPVPRNVLIPHTKGFVSSVSHMRSFVPAIYDVTVAFPKTSPPPTMLKLFEGQSVELHVHIKRHAMKDLPESDDAVAQWCRDKFVEKDALLDKHNSEDTFSGQEVHHVGRPIKALLVVISWVVVIIFGALKFLLWSSLLSSWKGKAFSVIGLGIVAGIVTLLMHILILSSQAEGSNPVKAAPAKLKTELSSSKKVTNKEN ChookLPAAT2aSEQ ID NO: 30  LSLLFFVSGLIVNLVQAVCFVLIRPLSKNTYRRINRVVAELLWLELVWLIDWWAGVKIKVFTDHETFNLMGKEHALVVCNHKSDIDWLVGWVLAQRSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYLFLERSWAKDESTLKSGLKRLKDYPLPFWLALFVEGTRFTQAKLLAAQQYAASSGLPVPRNVLIPRTKGFVSSVSHMRSFVPAIYDVTVAIPKTSVPPTMLRIFKGQSSVLHVHLKRHLMKDLPESDDAVAQWCRDIFVEKDALLDKHNAEDTFSGQELQDIGRPIKSLLVVISWAVLVIFGAVKFLQWSSLLSSWKGLAFSGIGLGIVTLLMHILILFSQSERSTPAKVAPAKPKNEGESSKTEMEKEH ChookLPAAT2b SEQ ID NO: 31QIKVFTDHETFNLMGKEHALVVCNHKSDIDWLVGWVLAQWSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYLFLERSWAKDESTLKSGLKRLKDYPLPFWLALFVEGTRFTQAKLLAAQQYAASSGLPVPRNVLIPRTKGFVSSVSHMRSFVPAIYDVTVAIPKTSVPPTMLRIFKGQSSVLHVHLKRHLMKDLPESDDAVAQWCRDIFVEKDALLDKHNAEDTFSGQELQDIGRPIKSLLVVISWAVLVIFGAVKFLQWSSLLSSWKGLAFSGIGLGIVTLLMHILILFSQSERSTPAKVAPAKLKKEGESSKPETDKQN ChookLPAAT3a SEQ ID NO: 32LSLLFFVSGLIVNLVQAVCFVLIRPLLKNTYRRINRVVAELLWLELVWLIDWWAGIKIKVFTDHETFHLMGKEHALVICNHKSDIDWLVGWVLAQRSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYLFLERNWAKDESTLKSGLNRLKDYPLPFWLALFVEGTRFTRAKLLAAQQYAASSGLPVPRNVLIPRTKGFVSSVSQMRSFVPAIYDVTVAIPKTSPPPTLLRMFKGQSSVLHVHLKRHLMNDLPESDDAVAQWCRDIFVEKDALLDKHNAEDTFSGQELQDTGRPIKSLLVVISWATLVVFGAVKFLQWSSLLSSWKGLAFSGIGLGIITLLMHILILFSQSERSTPAKVAPAKPKNEGESSKTEMEKEH ChookLPAAT3b SEQ ID NO: 33LSLLFFVSGLIVNLVQAVCFVLIRPLLKNTYRRINRVVAELLWLELVWLIDWWAGIKIKVFTDHETFHLMGKEHALVICNHKSDIDWLVGWVLAQRSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYLFLERNWAKDESTLKSGLNRLKDYPLPFWLALFVEGTRFTRAKLLAAQQYAASSGLPVPRNVLIPRTKGFVSSVSQMRSFVPAIYDVTVAIPKTSPPPTLLRMFKGQSSVLHVHLKRHLMNDLPESDDAVAQWCRDIFVEKDALLDKHNAEDTFSGQELQDIGRPIKSLLVVISWAVLEIFGAVKFLQWSSLLSSWKGLAFSGIGLGIVTLLMHILILFSQSERSTPAKVAPAKPKKEGESSKPETDKEN CigneaLPAAT1 SEQ ID NO: 34MAIAAAAVIFLFGLLFFASGIIINLFQALCFVLIWPLSKNVYRRINRVFAELLLMDLLCLFHWWAGAKIKLFTDPETFRLMGMEHALVIMNHKTDLDWMVGWILGQHLGCLGSILSIAKKSTKFIPVLGWSVWFSEYLFLERSWAKDKSTLKSHMEKLKDYPLPFWLVIFVEGTRFTRTKLLAAQQYAASSGLPVPRNVLIPHTKGFVSCVSNMRSFVPAVYDVTVAFPKSSPPPTMLKLFEGQSIVLHVHIKRHALKDLPESDDAVAQWCRDKFVEKDALLDKHNAEDTFSGQEVHHIGRPIKSLLVVIAWVVVIIFGALKFLQWSSLLSTWKGKAFSVIGLGIATLLMHMLILSSQAERSNPAKVAK CigneaLPAAT2 SEQ ID NO: 35MAIAAAAVIFLFGLLFFASGIIINLFQALCFVLIWPLSKNVYRRINRVFAELLLMDLLCLFHWWAGAKIKLFTDPETFRLMGMEHALVIMNHKTDLDWMVGWILGQHLGCLGSILSIAKKSTKFIPVLGWSVWFSEYLFLERSWAKDESTLKSGLNRLKDYPLPFWLALFVEGTRFTRAKLLAAQQYAASSGLPVPKNVLIPRTKGFVSSVSHMRSFVPAIYDVTVAIPKTSAPPTLLRMFKGQSSVLHVHLKRHLMKDLPESDDAVAQWCRDIFVEKDALLDKHNAEDTFSGQELHDIGRPVKSLLVVISWAMLVVFGAVKFLQWSSLLSSWKGLAFSGIGLGIITLLMHILILFSQSERSTPAKVAPAKQKNNEGESSKTEMEKEH DeLPAAT1 SEQ ID NO: 36SGLVVNLIQAFFFVLVRPFSKNAYRKINRVVAELLWLELIWLIDWWAGVKIQLYTDPETFKLMGKEHALVICNHKSDIDWLVGWILAQRSGCLGSALAVMKKSSKFLPVIGWSMWFSEYLFLERSWAKDENTLKSGFQRLRDFPHAFWLALFVEGTRFTQAKLLAAQEYASSMGLPAPRNVLIPRTKGFVTAVTHMRPFVPAVYDVTLAIPKTSPPPTMLRLFKGQSSVVHIHLKRHLMSDLPKSDDSVAQWCKDAFVVKDNLLDKHKENDSFGDGVLQDTGRPLNSLVVVISWACLLIFGALKFFQWSSILSSWKGLAFSAVGLGIVTVLMQILIQFSQSER SNRPMPSKHAK DeLPAAT2SEQ ID NO: 37  MAIPTAAYVVPLGAIFFFSGLLVNLIQAFFFITVWPLSKKTYIRINKVIVELLWLEFVWLADWWAGLKIEVYADAETFQLMGKEHALVICNHKSDIDWLVGWILAQRAGCLGSSFAVTKKSARYLPVVGWSIWFSGAIFLERSWEKDENTLKAGFQRLREFPCAFWLGLFVEGTRFTQAKLLAAQEYASTMGLPFPRNVLIPRTKGFIAAVNHMREFVPAIYDLTFAFPKDSPPPTMLRLLKGQPSVVHVHIKRHLMKDLPEKNEAVAQWCKDVFLVKDKLLDKHKDDGSFGDGELHEIGRPLKSLVVVTTWACLLILGTLKFLLWSSLLSSWKGLIFSATGLAVLTVLMQFLIQSTQSERSNPASLSK CerLPAAT1a SEQ ID NO: 38LGLLFFISGLAVNLIQAVCFVFLRPLSKNTYRKINRVLAELLWLQLVWLVDWWAGVKIKVFADRESFNLMGKEHALVICNHKSDIDWLVGWVLAQRSGCLGSSLAVMKKSSKFLPVIGWSMWFSEYLFLERSWAKDESTLKEGLRRLKDFPRPFWLALFVEGTRFTQAKLLAAQEYATSQGLPVPRNVLIPRTKVHVHVKRHLMKELPETDEAVAQWCKDLFVEKDKLLDKHVAEDTFSDQPLQDIGRPVKPLLVVSSWACLVAYGALKFLQWSSLLSSWKGIAVSAVALAIVT ILMQIMILFSQSERSIPAKVACerLPAAT1b SEQ ID NO: 39LGLLFFISGLAVNLIQAVCFVFLRPLSKNTYRKINRVLAELLWLQLVWLVDWWAGVKIKVFADRESFNLMGKEHALVICNHKSDIDWLVGWVLAQRSGCLGSSLAVMKKSSKFLPVIGWSMWFSEYLFLERSWAKDESTLKEGLRRLKDFPRPFWLALFVEGTRFTQAKLLAAQEYATSQGLPVPRNVLIPRTKGFVSAVSHMRSFVPAVYDMTVAIPKSSPSPTMLRLFKGQSSVVHVHVKRHLMKELPETDEAVAQWCKDLFVEKDKLLDKHVAEDTFSDQPLQDIGRPVKPLLVVSSWACLVAYGALKFLQWSSLLSSWKGIAVSAVALAIVTILMQIMI LFSQSERSIPTKVACerLPAAT2a SEQ ID NO: 40MAIAAAAVVFLFGLLFFTSGLIINLAQAVCFVLIWPLSKNAYRRINRVFAELLLLELLWLFHWRAGAKLKLFADPETFRLFGKEHALVICNHRTDLDWMVGWVLGQHFGCLGSILSVAKKSTKFLPVLGWSMWFSEYLFLERSWAKDKSTLKSHTERLKDYPLPFWLGIFVEGTRFTRAKLLAAQQYAASSGLPVPRNVLIPHTKLHVHIKRYAMKDLPESDDAVAQWCRDIYVEKDAFLDKHNAEDTFSGQEVHHIGRPIKSLLVVISWVVVIIFGALKFLRWSSLLSSWKGKAFSVIGLGIVTLLVNILILSSQAERSNPAKVAPAKLKTELSPSKKVTNK EN CerLPAAT2bSEQ ID NO: 41 MAIAAAAVVFLFGLLFFTSGLIINLAQAVCFVLIWPLSKNAYRRINRVFAELLLLELLWLFHWRAGAKLKLFADPETFRLFGKEHALVICNHRTDLDWMVGWVLGQHFGCLGSILSVAKKSTKFLPVLGWSMWFSEYLFLERSWAKDKSTLKSHTERLKDYPLPFWLGIFVEGTRFTRAKLLAAQQYAASSGLPVPRNVLIPHTKGFVSSMSHMRSFVPAVYDLTVAFPKTSPPPTLLKLFEGQSVVLHVHIKRYAMKDLPESDDAVAQWCRDIYVEKDAFLDKHNAEDTFSGQEVHHIGRPIKSLLVVISWVVVIIFGALKFLRWSSLLSSWKGKAFSVIGLGIVTLLVNILILSSQAERSNPAKVAPAKLKTELSPSKKVTNKEN BrLPAAT1a SEQ ID NO: 42AAAVIVPLGILFFISGLVVNLLQAICYVLIRPLSKNTYRKINRVVAETLWLELVWIVDWWAGVKIQVFADNETFNRMGKEHALVVCNHRSDIDWLVGWILAQRSGCLGSALAVMKKSSKFLPVIGWSMWFSEYLFLERNWAKDESTLKSGLQRLNDFPRPFWLALFVEGTRFTEAKLKAAQEYAASSELPVPRNVLIPRTKGFVSAVSNMRSFVPAIYDMTVAIPKTSPPPTMLRLFKGQPSVVHVHIKCHSMKDLPESDDAIAQWCRDQFVAKDALLDKHIAADTFPGQQEQNIGRPIKSLAVVLSWSCLLILGAMKFLHWSNLFSSWKGIAFSALGLGIITLCMQILIRSSQSERSTPAKVVPAKPKDNHNDSGSSSQTE BrLPAAT1b SEQ ID NO: 43AAAVIVPLGILFFISGLVVNLLQAVCYVLVRPMSKNTYRKINRVVAETLWLELVWIVDWWAGVKIQVFADDETFNRMGKEHALVVCNHRSDIDWLVGWILAQRSGCLGSALAVMKKSSKFLPVIGWSMWFSEYLFLERNWAKDESTLKSGLQRLNDFPRPFWLALFVEGTRFTEAKLKAAQEYAASSELPVPRNVLIPRTKGFVSAVSNMRSFVPAIYDMTVAIPKTSPPPTMLRLFKGQPSVVHVHIKCHSMKDLPESDDAIAQWCRDQFVAKDALLDKHIAADTFPGQQEQNIGRPIKSLAVVLSWSCLLILGAMKFLHWSNLFSSWKGIAFSALGLGIITLCMQILIRSSQSERSTPAKVVPAKPKDNHNDSGSSSQTE BrLPAAT1e SEQ ID NO: 44 MAIAAAVIVPLGLLFFISGLLMNLLQAICYVLVRPLSKNTYRKINRVVAETLWLELVWIVDWWAGVKIKVFADNETFSRMGKEHALVVCNHRSDIDWLVGWILAQRSGCLGSALAVMKKSSKFLPVIGWSMWFSEYLFLERNWAKDESTLKSGLQRLNDFPRPFWLALFVEGTRFTEAKLKAAQEYAASSELPVPRNVLIPRTKGFVSAVSNMRSFVPAIYDMTVAIPKTSPPPTMLRLFKGQPSVVHVHIKCHSMKDLPESDDAIAQWCRDQFVAKDALLDKHIAADTFPGQQEQNIGRPIKSLAVVLSWSCLLILGAMKFLHWSNLFSSWKGIAFSALGLGIITLCMQILIRSSQSERSTPAKVVPAKPKDNHNDSGSSSQTE BjLPAAT1a SEQ ID NO: 45 INLVVAETLWLELVWIVDWWAGVKIQVFADDETFNRMGKEHALVVCNHRSDIDWLVGWILAQRSGCLGSALAVMKKSSKFLPVIGWSMWFSEYLFLERNWAKDESTLKSGLQRLNDFPRPFWLALFVEGTRFTEAKLKAAQEYAASSELPVPRNVLIPRTKGFVSAVSNMRSFVPAIYDMTVAIPKTSPPPTMLRLFKGQPSVVHVHIKCHSMKDLPESDDAIAQWCRDQFVAKDALLDKHIAADTFPGQKEQNIGRPIKSLAVSLIKTFPWLHPHQLTNIFVLFQVVVSWACLLTLGAMKFLHWSNLFSSWKGIALSAFGLGIITLCMQILIRSSQSERSTP AKVAPAKPK BjLPAAT1bSEQ ID NO: 46  INLVVAETLWLELVWIVDWWAGVKIQVFADDETFNRMGKEHALVVCNHRSDIDWLVGWILAQRSGCLGSALAVMKKSSKFLPVIGWSMWFSEYLFLERNWAKDESTLKSGLQRLNDFPRPFWLALFVEGTRFTEAKLKAAQEYAASSELPVPRNVLIPRTKGFVSAVSNMRSFVPAIYDMTVAIPKTSPPPTMLRLFKGQPSVVHVHIKCHSMKDLPEPEDEIAQWCRDQFVAKDALLDKHIAADTFPGQKEQNIGRPIKSLAVVVSWACLLTLGAMKFLHWSNLFSSWKGIALSAFGLGIITLCMQILIRSSQSERSTPAKVAPAKPK BjLPAAT1e SEQ ID NO: 47 INLVVAETLWLELVWIVDWWAGVKIQVFADDETFNRMGKEHALVVCNHRSDIDWLVGWILAQRSGCLGSALAVMKKSSKFLPVIGWSMWFSEYLFLERNWAKDESTLKSGLQRLNDFPRPFWLALFVEGTRFTEAKLKAAQEYAASSELPVPRNVLIPRTKGFVSAVSNMRSFVPAIYDMTVAIPKTSPPPTMLRLFKGQPSVVHVHIKCHSMKDLPESDDAIAQWCRDQFVAKDALLDKHIAADTFPGQQEQNIGRPIKSLAVVLSWSCLLILGAMKFLHWSNLFSSWKGIAFSALGLGIITLCMQILIRSSQSERSTPAKVVPAKPKDNHNDSGSSSQTE BjLPAAT1dSEQ ID NO: 48  INLVVAETLWLELVWIVDWWAGVKIQVFADDETFNRMGKEHALVVCNHRSDIDWLVGWILAQRSGCLGSALAVMKKSSKFLPVIGWSMWFSEYLFLERNWAKDESTLKSGLQRLNDFPRPFWLALFVEGTRFTEAKLKAAQEYAASSELPVPRNVLIPRTKGFVSAVSNMRSFVPAIYDMTVAIPKTSPPPTMLRLFKGQPSVVHVHIKCHSMKDLPESDDAIAQWCRDQFVAKDALLDKHIAAD TFPGQQEQNIGRPIKSLAVSLSCeLPAAT1a SEQ ID NO: 49MAIGVAAIVVPLGLLFILSGLMVNLIQAICFILVRPLSKNMYRRVNRVVVELLWLELIWLIDWWGGVKVDVYADSETFQSLGKEHALVVSNHRSDIDWLVGWVLAQRSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYVFLERSWAKDESTLKSGLRRLKDFPRPFWLALFVEGTRFTQAKLLAAREYAASTGLPIPRNVLIPRTKGFVSAVSNMRSFVPAIYDVTVAIPKTQPSPTMLRIFNRQPSVVHVHIKRHSMNQLPQTDEGVGQWCKDIFVAKDALLDRHLAE CcLPAAT1b SEQ ID NO: 50 MAIGVAAIVVPLGLLFILSGLMVNLIQAICFILVRPLSKNMYRRVNRVVVELLWLELIWLIDWWGGVKVDVYADSETFQSLGKEHALVVSNHRSDIDWLVGWVLAQRSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYVFLERSWAKDESTLKSGLRRLKDFPRPFWLALFVEGTRFTQAKLLAAREYAASTGLPIPRNVLIPRTKGFVSAVSNMRSFVPAIYDVTVAIPKTQPSPTMLRIFNRQPSVVHVHIKRHSMNQLPQTDEGVAQWCKDIFVAKDALLDRHLAEGKFDEKEFKRIRRPIKSLLVISSWSFLLMFGVFKFLKWSALLSTWKGVAVSTTVLLLVTVVMYMFILFSQSERSSPRKVAPSGPENG UcLPAAT1a SEQ ID NO: 51 MAIGVAAIVVPLGLLFILSGLIINLIQAICFILVRPLSKNMYRKVNRVVVELLWLELIWLIDWWGGVKVDVYADSETFQSLGKEHALVVSNHRSDIDWLVGWVLAQRSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYVFLERSWAKDESTLKSGLQRLKDFPRPFWLALFVEGTRFTQAKLLAAQEYAASTGLPIPRNVLIPRTKGFVSAVSNMRSFVPAIYDVTVAIPKTQPSPTMLRIFNRQPSVVHVHIKRHSMNQLPQTDEGVAQWCKDIFVAKDALLDRHLAEGKFDEKEFKLIRRPIKSLLVISSWSFLLMFGVFKFLKWSALLSTWKGVAVSTAVLLLVTVVMYMFILFSQSERSSPRKVAPIGPENG UcLPAAT1b SEQ ID NO: 52 MAIGVAAIVVPLGLLFILSGLIINLIQAICFILVRPLSKNMYRKVNRVVVELLWLELIWLIDWWGGVKVDVYADSETFQSLGKEHALVVSNHRSDIDWLVGWVLAQRSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYVFLERSWAKDESTLKSGLQRLKDFPRPFWLALFVEGTRFTQAKLLAAQEYAASTGLPIPRNVLIPRTKGFVSAVSNMRSFVPAIYDVTVAIPKTQPSPTMLRIFNRQPSVVHVHIKRHSMNQLPQTDEGVAQWCKDIFVAKDALLDRHLAE LdLPAAT1 SEQ ID NO: 53 SLLFFMSGLVVNFIQAVFYVLVRPISKNTYRRINTLVAELLWLELVWVIDWWAGVKVQLYTDTESFRLMGKEHALLICNHRSDIDWLIGWVLAQRCGCLSSSIAVMKKSSKFLPVIGWSMWFSEYLFLERNWAKDENTLKSGLQRLNDFPKPFWLALFVEGTRFTKAKLLAAQEYAASAGLPVPRNVLIPRTKGFVSAVSNMRSFVPAIYDLTVAIPKTTEQPTMLRLFRGKSSVVHVHLKRHLMKDLPKTDDGVAQWCKDQFISKDALLDKHVAEDTFSGLEVQDIGRPMKSLVVVVSWMCLLCLGLVKFLQWSALLSSWKGMMITTFVLGIVTVLMHILIR SSQSEHSTPAKCaequLPAAT1a SEQ ID NO: 54 QRSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYLFLERSWAKDESTLKSGLKRLKDYPLPFWLALFVEGTRFTQAKLLAAQQYAASSGLPVPRNVLIPRPTKGFVSSVSHMRSFVAIYDVTVAIPKMSTPPTMLRIFKGQSSVLHVHLKRHLMKDLPESDDAVAQWCRDIFVEKDALLDKHNAEDTFSGQELQDIGRPVKSLLVVISWAVLVIFGAVKFLQWSSLLSSWKGLAFSGIGLGIVTLLMHILILFSQSERSTPAKVAPAKPKKEGESSKTETEKEN CaequLPAAT1b SEQ ID NO: 55 DWWAGVKIKVFTDHETLSLMGKEHALVISNHKSDIDWLVGWVLAQRSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYLFLERSWAKDESTLKSGLKRLKDYPLPFWLALFVEGTRFTQAKLLAAQQYAASSGLPVPRNVLIPRTKGFVSSVSHMRSFVPAIYDVTVAIPKMSTPPTMLRIFKGQSSVLHVHLKRHLMKDLPESDDAVAQWCRDIFVEKDALLDKHN AEDTFSGQELQDIGRPVKSLLV CaequLPAAT1eSEQ ID NO: 56 DWWAGVKIKVFTDHETLSLMGKEHALVISNHKSDIDWLVGWVLAQRSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYLFLERSWAKDESTLKSGLKRLKDYPLPFWLALFVEGTRFTQAKLLAAQQYAASSGLPVPRNVLIPRTKGFVSSVSHMRSFVPAIYDVTVAIPKMSTPPTMLRIFKGQSSVLHVHLKRHLMKDLPESDDAVAQWCRDIFVEKDALLDKHNAEDTFSGQELQDIGRPVKSLLVVISWAVLVIFGAVKFLQWSSLLSSWKGLAFSGIGLGIVTLLMHILILFSQSERSTPAKVAPAKPKKEGESSKTETEKEN CaequLPAAT1d SEQ ID NO: 57QRSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYLFLERSWAKDESTLKSGLKRLKDYPLPFWLALFVEGTRFTQAKLLAAQQYAASSGLPVPRNVLIPRTKGFVSSVSHMRSFVPAIYDVTVAIPKMSTPPTMLRIFKGQSSVLHVHLKRHLMKDLPESDDAVAQWCRDIFVEKDALLDKHNAEDTFSGQELQDIGR PVKSLLV CglutLPAAT1aSEQ ID NO: 58  LSLLFFVSGLFVNLVQAVCFVLIRPFSKNTYRRINRVVAELLWLELVWLIDWWAGVKIKVFTDHETLSLMGKEHALVISNHKSDIDWLVGWVLAQRSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYLFLERSWAKDESTLKSGLKRLKDYPLPFWLALFVEGTRFTQAKLLAAQQYAASSGLPVPRNVLIPRTKGFVSSVSHMRSFVPAIYDVTVAIPKMSTPPTMLRIFKGQSSVLHVHLKRHLMKDLPESDDAVAQWCRDIFVEKDALLDKHNAEDTFSGQELQDIGRPVKSLLVVISWAVLVIFGAVKFLQWSSLLSSWKGLAFSGIGLGIVTLLMHILILFSQSERSTPAKVAPAKPKKEGESSKTETEKEN CglutLPAAT1b SEQ ID NO: 59QAVCFVLIRPFSKNTYRRINRVVAELLWLELVWLIDWWAGVKIKVFTDHETLSLMGKEHALVISNHKSDIDWLVGWVLAQRSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYLFLERSWAKDESTLKSGLKRLKDYPLPFWLALFVEGTRFTQAKLLAAQQYAASSGLPVPRNVLIPRTKGFVSSVSHMRSFVPAIYDVTVAIPKMSTPPTMLRIFKGQSSVLHVHLKRHLMKDLPESDDAVAQWCRDIFVEKDALLDKHNAEDTFSGQELQDIGRPVKSLLVVISWAVLVIFGAVKFLQWSSLLSSWKGLAFSGIGLGIVTLLMHILILFSQSERSTPAKVAP AKPKKEGESSKTETEKENCprLPAAT1 SEQ ID NO: 60MAIAAAAVVFLFGLLFFTSGLIINLAQAVCFVLIWPLSKNAYRRINRVFAELLLLELLWLFHWRAGAKLKLFADPETFRLFGKEHALVICNHRTDLDWMVGWVLGQHFGCLGSILSVAKKSTKFLPVLGWSMWFSEYLFLERSWAKDKSTLKSHTERLKDYPLPFWLGIFVEGTRFTRAKLLAAQQYAASSGLPVPRNVLIPHTKGFVSSMSHMRSFVPAVYDLTVAFPKTSPPPTLLKLFEGQSVVLHVHIKRYAMKDLPESDDAVAQWCRDIYVEKDAFLDKHNAEDTFSGQEVHHIGRPIKSLLVVISWVVVIIFGALKFLRWSSLLSSWKGKAFSVIGLGIVTLLVNILILSSQAERSNPAKVVPAKLKTELSPSKKVTNKEN ChsLPAAT1 SEQ ID NO: 61MAIPSAAVVFLFGLLFFASGLIINLVQAVCFVLIWPLSKNTCRRINIVFQDMLLSELLWLFHWRAGAKLKFFTDPETYRHMGKEHALVITNHRTDLDWMIGWVLGEHLGCLGSILSVVKKSTKFLPVLGWSMWFSEYLFLERNWAKDKSTFKSHIERLEDFPQPFWFGIFVEGTRFTRAKLLAAQQYAASSGLPVPRNVLIPHTKGFVSSVSHMRSFVPAVYETTMTFPKTSPPPTLLKLFEGQPLVLHIHMKRHAMKDIPESDDAVAQWCRDKFVEKDALLDKHNAEDTFGGLEVHIGRSIKSLMVVICWVVVIIFGALKFLQWSSLLSSWKGIAFIGIGLGIVNLLVHVLILSSQAERSAPTKVAPAKLKTKLLSSKKITNKEN ChsLPAAT2 SEQ ID NO: 62 MAIPSAAVVFLFGLLFFASGLIINLVQAVCFVLIWPLSKNTCRRINIVFQDMLLSELLWLFHWRAGAKLKFFTDPETYRHMGKEHALVITNHRTDLDWMIGWVLGEHLGCLGSILSVVKKSTKFLPVLGWSMWFSEYLFLERNWAKDKSTFKSHIERLEDFPQPFWFGIFVEGTRFTRAKLLAAQQYAASSGLPVPRNVLIPRTKGFVSSVSHMRSFVPAIYDVTVAIPKTSPPPTMLRMFKGQSSVLHVHLKRHLMKDLPESDDAVAQWCRDIFVEKDALLDKHNAEDTFSGQELQDIGRPIKSLVVVISWAALVVFGAVKFLQWSSLLSSWKGLAFSGIGLGIITLLMHILILFSQSERSTPAKVAPAKPKREGESSKTEMDKEN CcaleLPAAT1a SEQ ID NO: 63MAIPAAAVVFLFGLLFFPSGLIINLFQAVCFVLIWPFSRNTCRRINIVFQEMLLSELLWLFHWRAGAKLKLFADPETYRHMGKEHALLITNHRTDLDWMIGWALGQHLGCLGSILSVVKKSTKFLPSHIERLEDFPQPFWMAIFVEGTRFTRAKLLAAQQYAASSGLPVPRNVLIPRTKGFVSCVSHMRSFVPAVYETTMTFPKTSPPPTLLKLFEGQPIVLHVHMKRHAMKDIPESDEAVAQWCRDKFVEKDSLLDKHNAGDTFSCQEIHIGRPIKSLMVVISWVVVIIFGALKFLQWSSLLSSWKGIAFSGIGLGIVTLLVHILILSSQAERSTPAKVAPA KLKTELSSSTKVTNKENCcaleLPAAT1b SEQ ID NO: 64 MAIPAAAVVFLFGLLFFPSGLIINLFQAVCFVLIWPFSRNTCRRINIVFQEMLLSELLWLFHWRAGAKLKLFADPETYRHMGKEHALLITNHRTDLDWMIGWALGQHLGCLGSILSVVKKSTKFLPVLGWSMWFSEYLFLERNWAKDKSTFKSHIERLEDFPQPFWMAIFVEGTRFTRAKLLAAQQYAASSGLPVPRNVLIPRTKGFVSCVSHMRSFVPAVYETTMTFPKTSPPPTLLKLFEGQPIVLHVHMKRHAMKDIPESDEAVAQWCRDKFVEKDSLLDKHNAGDTFSCQEIHIGRPIKSLMVVISWVVVIIFGALKFLQWSSLLSSWKGIAFSGIGLGIVTLLVHILILSSQAERSTPAKVAPAKLKTELSSSTKVTNKEN CcaleLPAAT2 SEQ ID NO: 65LSLLFFVSGLIVNLVQAVCFVLIRPLSKNTYRRINRVVAELLWLELVWLIDWWAGVKIKVFTDHETFRLMGTEHALVISNHKSDIDWLVGWVLAQRSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYLFLERSWAKDESTLKSGLNRLKDYPLPFWLALFVEGTRFTRAKLLAAQQYAASSGLPVPRNVLIPRTKGFVSSVSHMRSFVPAIYDVTVAIPKTSPPPTMLRMFKGQSSVLHVHLKRHLMKDLPESDDAVAQWCRDIFVEKDALLDKHNAEDTFSGQELQDIGRPIKSLVVVISWAALVVFGAVKFLQWSSLLSSWKGLAFSGIALGIITLLMHILILFSQSERSTPAKVAPAKPKKEGESSKTETDKEN ChtLPAAT1a SEQ ID NO: 66MAIPAAAVIFLFSILFFASGLIINLVQAVCFVLIWPLSKNTCRRINLVFQEMLLSELLGLFHWRAGAKLKLYTDPETYPLLGKEHALLMINHRTDLDWMIGWVLGQHLGCLGSILSVVKKSTKFLPVLGWSMWFSEYLFLERNWAKDKSTFKSHIERLEDFPQPFWMAIFVEGTRFTRAKLLAAQQYAASSGLPVPRNVLIPHTKGFVSTVSHMRSFVPAVYDTTLTFPKTSPPPTLLNLFAGQPIVLHIHIKRHAMKDIPESDDAVAQWCRDKFVEKDALLDKHNAEDAFSDQEFPISRSIKSLMVVISWVMVIIFGALKFLQWSSLLSSWKGKAFSVIAVGIVTLLMHMSILSSQAERSNPAKVALPKLKTELPSSKKVLNKEN ChtLPAAT1b SEQ ID NO: 67MAIPAAAVIFLFSILFFASGLIINLVQAVCFVLIWPLSKNTCRRINLVFQEMLLSELLGLFHWRAGAKLKLYTDPETYPLLGKEHALLMINHRTDLDWMIGWVLGQHLGCLGSILSVVKKSTKFLPVLGWSMWFSEYLFLERNWAKDKSTFKSHIERLEDFPQPFWMAIFVEGTRFTRAKLLAAQQYAASSGLPVPRNVLIPHTKGFVSTVSHMRSFVPAVYDTTLTFPKTSPPPTLLNLFAGQPIVLHIHIKRHAMKDIPESDDAVAQWCRDKFVEKDALLDKHNAEDAFSDQEFPISRSIKSLMVVISWVMVIIFGALKFLQWSSLLSSWKGIAFSGIGLGIVTLLMHILILSSQAERSTPAKVAQAKVKTELPSSTKVTNKGN CwLPAAT1 SEQ ID NO: 68MAIPAAAVIFLFGILFFASGLIINLVQAVCFVLIWPLSKNTCRRINLVFQEMLLSELLWLFHWRAGAELKLFTDPETYRLLGKEHALVMTNHRTDLDWMIGWVTGQHLGCLGSILSIAKKSTKFLPVLGWSMWFSEYLFLERNWAKDKSTFKSHIERLEDFPQPFWMAIFVEGTRFTRAKLLAAQQYAASSGLPVPRNVLIPHTKGFVSSVCHMRSFVPAVYDTTLTFPKNSPPPTLLNLFAGQPIVLHIHIKRHAMKDMPKSDDAVAQWCRDKFVKKDALLDKHNTEDTFSDQEFPIGRPIKSLMVVISWVVVIIFGTLKFLQWSSLLSSWKGIAFSGIGLGIVTLLVHILILSSQAERSTPPKVAPAKLKTELSSTTKVINKGN CwLPAAT2b SEQ ID NO: 69LGLLFFVSGLIVNLVQAVCFVLIRPLSKNTYRRLNRVVAELLWLELVWLIDWWAGVKIKVFTDHETFHLMGKEHALVICNHKSDIDWLVGWVLAQRSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYLFLERSWAKDESTLKSGLNRLKDYPLPFWLALFVEGTRFTRAKLLAAQQYAASSGLPVPRNVLIPRTKGFRVSSVSHMRSFVPAIYDVTVAIPKTSPPPTMLMFKGQSSVDALLDKHNADDTFSGQELHDIGRPIKSLLVVISWAVLVVFGAVKFLQWSSLLSSWKGIAFSGIGLGIVTLLVHILILSSQAERSTSAKVAQAKVKTELSSSKKVKNK GN CwLPAAT2aSEQ ID NO: 70 LGLLFFVSGLIVNLVQAVCFVLIRPLSKNTYRRLNRVVAELLWLELVWLIDWWAGVKIKVFTDHETFHLMGKEHALVICNHKSDIDWLVGWVLAQRSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYLFLERSWAKDESTLKSGLNRLKDYPLPFWLALFVEGTRFTRAKLLAAQQYAASSGLPVPRNVLIPRTKGFVSSVSHMRSFVPAIYDVTVAIPKTSPPPTMLRMFKGQSSVLHVHLKRHLMKDLPESDDAVAQWCRDIFVEKDVLLDKHNAEDTFSGQELQDIGRPVKSLLVVISWTLLVIFGAVKFLQWSSLLSSWKGLAFSGIGLGIVTLLMHILILFSQSERSTPAKVAPAKPKKEGESSKMETDKEN CgLPAAT1a SEQ ID NO: 71LAGWMGSSSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYLFLERSWAKDESTLKSGLNRLKDYPLPFWLALFVEGTRFTRAKLLAAQQYAASLGLPVPRNVLIPRTKGFVSSVSHMRSFVPAIYDVTVAIPKTSPPPTMIRMFKGQSSVLHVHLKRHVMKDLPESDDAVAQWCRDIFVEKDALLDKHNAEDTFSGQELQDTGRPIKSLLVVISWAVLEVFGAVKFLQWSSLLSSWKGLAFSGIGLGIITLLMHILILFSQSERSTPAKVAPAKPKNEGESSKAEMEKEK CgLPAAT1b SEQ ID NO: 72LAGWMGSSSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYLFLERSWAKDESTLKSGLNRLKDYPLPFWLALFVEGTRFTRAKLLAAQQYAASLGLPVPRNVLIPRTKGFVSSVSHMRSFVPAIYDVTVAIPKTSPPPTMIRMFKGQSSVLHVHLKRHVMKDLPESDDAVAQWCRDIFVEKDALLDKHNAEDTFSGQELQDTGRPIKSLLVRCFLVLSLIYLNGIMLKLRGPCLQVVISWAVLEVFGAVKFLQWSSLLSSWKGLAFSGIGLGIITLLMHILILFSQSERSTPAKVA PAKPKNEGESSKAEMEKEKCgLPAAT1c SEQ ID NO: 73LAGWMGSSSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYLFLERSWAKDESTLKSGLNRLKDYPLPFWLALFVEGTRFTRAKLLAAQQYAASLGLPVPRNVLIPRTKGFVSSVSHMRSFVPAIYDVTVAIPKTSPPPTMIRMFKGQSSVLHVHLKRHVMKDLPESDDAVAQWCRDIFVEKDALLDKHNAEDTFSGQELQDTGRPIKSLLVVTSWAVLVISGAVKFLQWSSLLSSWKGLAFSGIGLGIVTLLMHILILFSQSERSTPAKVAPAKPKKEGESSKTEKDKEN CpalLPAAT1 SEQ ID NO: 74LGLLFFVSGLIVNLVQAVCFVLIRPLSKNTYRRINRVVAELLWLELVWLIDWWAGVKIKVFTDHETLSLMGKEHALVICNHKSDIDWLVGWVLAQRSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYLFLERSWAKDENTLKSGLNRLKDYPLPFWLALFVEGTRFTRAKLLAAQQYATSSGLPVPRNVLIPRTKGFVSSVSHMRSFVPAIYDVTVAIPKTSPPPTMLRMFKGQSSVLHVHLKRHLMKDLPESDDAVAQWCRDIFVEKDALLDKHNAEDTFSGQELQDTGRPIKSLLVVISWAVLVIFGAVKFLQWSSLLSSWKGLAFSGVGLGIITLLMHILILFSQSERSTPAKVAPAKPKKDGESSKTEIEKEN CaLPAAT1 SEQ ID NO: 75MAIAAAAVIVPVSLLFFVSGLIVNLVQAVCFVLIRPLFKNTYRRINRVVAELLWLELVWLIDWWAGVKIKVFTDHETFHLMGKEHALVICNHKSDIDWLVGWVLAQRSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYLFLERNWAKDESTLKSGLNRLKDYPLPFWLALFVEGTRFTRAKLLAAQQYAASSGLPVPRNVLIPRTKGFVSSVSHMRSFVPAIYDVTVAIPKTSPPPTLLRMFKGQSSVLHVHLKRHQMNDLPESDDAVAQWCRDIFVEKDALLDKHNAEDTFSGQELQDTGRPIKSLLIVISWAVLVVFGAVKFLQWSSLLSSWKGLAFSGIGLGVITLLMHILILFSQSERSTPAKVAPAKPKIEGESSKTEMEKEH CaLPAAT3 SEQ ID NO: 76MTIASAAVVFLFGILLFTSGLIINLFQAFCSVLVWPLSKNAYRRINRVFAEFLPLEFLWLFHWWAGAKLKLFTDPETFRLMGKEHALVIINHKIELDWMVGWVLGQHLGCLGSILSVAKKSTKFLPVFGWSLWFSEYLFLERNWAKDKKTLKSHIERLKDYPLPFWLIIFVEGTRFTRTKLLAAQQYAASAGLPVPTRNVLIPHTKGFVSSVSHMRSFVPAIYDVTVAFPKSPPPTMLKLFEGHFVELHVHIKRHAMKDLPESEDAVAQWCRDKFVEKDALLDKHNAEDTFSGQEVHHVGRPIKSLLVVISWVVVIIFGALKFLQWSSLLSSWKGIAFSVIGLGTVALLMQILILSSQAERSIPAKETPANLKTELSSSKKVTNKEN SalLPAAT1 SEQ ID NO: 77MAIGAAAIVVPLGLLFMLSGLMVNLIQAICFILVRPLSKNMYRRVNRVVVELLWLELIWLIDWWGGVKVDVYADSETFQSLGKEHALVVSNHKSDIDWLVGWVLAQRSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYVFLERSWAKDESTLKSGLQRLKDFPRPFWLALFVEGTRFTQAKLLAAQEYAASTGLPIPRNVLIPRTKGFVSAVSNMRSFVPAIYDVTVAIPKTQPSPTMLRIFNRQPSVVHVRIKRHSMNQLPPTDEGVAQWCKDIFVAKDALLDRHLAEGKFDEKEFKRIRRPIKSLLVISSWSFLLLFGVFKFLKWSALLSTWKGVAVSTAVLLLVTVVMYMFILFSQSERSSPRKVAPSGPENG CleptLPAAT1 SEQ ID NO: 78 MAIPAAVVIFLFGLLFFSSGLIINLFQALCFVLIWPLSKNAYRRINRVFAELLLSELLCLFDWWAGAKLKLFTDPETFRLMGKEHALVIINHMTELDWMVGWVMGQHFGCLGSILSVAKKSTKFLPVLGWSMWFTEYLYIERSWDKDKSTLKSHIERLKDYPLPFWLVIFAEGTRFTRTKLLAAQQYAASSGLPVPRNVLIPRTKGFVSCVNHMRSFVPAVYDLTVAFPKTSPPPTLLNLFEGQSVVLHVHIKRHAMKDLPESDDAVAQWCRDKFVEKDALLDKHNAEDTFSSQEVHHTGSRPIKSLLVVISWVVVITFGALKFLQWSSWKGKAFSVIGLGIVTLLMHMLILSSQAERSKPAKVTQAKLKTELSISKKVTDKEN ClopLPAAT1 SEQ ID NO: 79MAIAAAAVIFLFGLLFFASGLIINLFQALCFVLIRPLSKNAYRRINRVFAELLLSELLCLFDWWAGAKLKLFTDPETLRLMGKEHALIIINHMTELDWMVGWVMGQHFGCLGSIISVAKKSTKFLPVLGWSMWFSEYLYLERSWAKDKSTLKSHIERLKDYPLPFWLVIFVEGTRFTRTKLLAAQEYAASSGLPVPRNVLIPRTKGFVSCVNHMRSFVPAVYDVTVAFPKTSPQPTLLNLFEGRSIVLHVHIKRHAMKDLPESDDAVAQWCRDKFVEKDALLDKHNAEDTFSGQEVHHTGRRPIKSLLVVMSWVVVTTFGALKFLQWSSWKGKAFSVIGLGIVTLLMHVLILSSQAERSNPAKVVQAELNTELSISKKVTNKGN CcrasLPAAT1a SEQ ID NO: 80MAIPAAAVIFLFGLIFFASGLIINLFQALCFVLIWPLWKNAYRRINRVFAELLLSELLCLFDWWAGAKLKLFTDPETFRLMGKEHALVIINHMTELDWMVGWVMGQHFGCLGSILSVAKKSTKFLPVLGWSMWFTEYLYIERSWDKDKSTLKSHIERLKDYPLPFWLVIFAEGTRFTRTKLLAAQQYAASSGLPVPRNVLIPRTKGFVSSVSHMRSFVPAIYDVTVAIPKTSPPPTLIRMFKGQSSVLHVHLKRHVMKDLPESDDAVAQWCRDIFVEKDALLDKHNAEDTFSGQELQDTGRPIKSLLVVISWAVLEVFGAVKFLQWSSLLSSWKGLAFSGIGLGIITLLMHILILFSQSERSTPAKVAPAKAK CcrasLPAAT1b SEQ ID NO: 81MAIPAAAVIFLFGLIFFASGLIINLFQALCFVLIWPLWKNAYRRINRVFAELLLSELLCLFDWWAGAKLKLFTDPETFRLMGKEHALVIINHMTELDWMVGWVMGQHFGCLGSILSVAKKSTKFLPVLGWSMWFTEYLYIERSWDKDGKSTLKSHIERLKDYPLPFWLVIFAETRFTRTKLLAAQQYAASSGLPVPRNVLIPRTKGFVSSVSHMRSFVPAIYDVTVAIPKTSPPPTLIRMFKGQSSVLHVHLKRHVMKDLPESDDAVAQWCRDIFVEKDALLDKHNAEDTFSGQELQDTGRPIKSLLVRCFLVLSLIYLNGIILKLCGLCLQVVISWAVLEVFGAVKFLQWSSLLSSWKGLAFSGIGLGIITLLMHILILFSQSERSTPAKV APAKAK CcrasLPAAT1cSEQ ID NO: 82 MAIPAAAVIFLFGLIFFASGLIINLFQALCFVLIWPLWKNAYRRINRVFAELLLSELLCLFDWWAGAKLKLFTDPETFRLMGKEHALVIINHMTELDWMVGWVMGQHFGCLGSILSVAKKSTKFLPVLGWSMWFTEYLYIERSWDKDKSTLKSHIERLKDYPLPFWLVIFAEGTRFTRTKLLAAQQYAASSGLPVPRNVLIPRTKGFVSSVSHMRSFVPAIYDVTVAIPKTSPPPTLIRMFKGQSSVLHVHLKRHVMKDLPESDDAVAQWCRDIFVEKDALLDKHNAEDTFSGQELQDTGRPIKSLLVVISWAVLEVFGAVKFLQWSSLLSSWKGLAFSGIGLGIITLLMHILILFSQSERSTPAKVAPAKAKMEGESSKTEMEMEK CcrasLPAAT1d SEQ ID NO: 83MAIPAAAVIFLFGLIFFASGLIINLFQALCFVLIWPLWKNAYRRINRVFAELLLSELLCLFDWWAGAKLKLFTDPETFRLMGKEHALVIINHMTELDWMVGWVMGQHFGCLGSILSVAKKSTKFLPVLGWSMWFTEYLYIERSWDKDKSTLKSHIERLKDYPLPFWLVIFAEGTRFTRTKLLAAQQYAASSGLPVPRNVLIPRTKGFVSSVSHMRSFVPAIYDVTVAIPKTSPPPTLIRMFKGQSSVLHVHLKRHVMKDLPESDDAVAQWCRDIFVEKDALLDKHNAEDTFSGQELQDTGRPIKSLLVRCFLVLSLIYLNGIILKLCGLCLQVVISWAVLEVFGAVKFLQWSSLLSSWKGLAFSGIGLGIITLLMHILILFSQSERSTPAKV APAKAKMEGESSKTEMEMEKCkoeLPAAT1 SEQ ID NO: 84 MAIAAAPVIFLFGLLFFASGLIINLFQAICFVLIWPLSKNAYRRINRVFAELLLSELLCLFDWWAGAKLKLFTDPETFRLMGKEHALVITNHKIDLDWMIGWILGQHFGCLGSVISIAKKSTKFLPIFGWSLWFSEYLFLERNWAKDKRTLKSHIERMKDYPLPLWLILFVEGTRFTRTKLLAAQQYAASSGLPVPRNVLIPHTKGFVSSVSHMRSFVPAIYDVTVAIPKTSPPPTLIRMFKGQSSVLHVHLKRHLMKDLPESDDAVAQWCRDIFVEKDALLDKHNAEDTFSGQELQETGRPIKSLLVVISWAVLEVYGAVKFLQWSSLLSSWKGLAFSGIGLGLITLLMHILILFSQSERSTPAKVAPAKPKKEGESSKTEMEKEK CkoeLPAAT2 SEQ ID NO: 85MHVLLEMVTFRFSSFFVFDNVQALCFVLIWPLSKSAYRKINRVFAELLLSELLCLFDWWAGAKLKLFTDPETFRLMGKEHALVITNHKIDLDWMIGWILGQHFGCLGSVISIAKKSTKFLPIFGWSLWFSEYLFLERNWAKDKRTLKSHIERMKDYPLPLWLILFVEGTRFTRTKLLAAQQYAASSGLPVPRNVLIPHTKGFVSSVSHMRSFVPAVYDVTVAFPKTSPPPTMLSLFEGQSVVLHVTHIKRHAMKDLPDSDDAVAQWCRDKFVEKDALLDKHNAEDFSGQEVHHVGRPIKSLLVVISWMVVIIFGALKFLQWSSLLSSWKGKAFSAIGLGIATLLMHVLVVFSQADRSNPAKVPPAKLNTELSSSKKVTNKEN

Example 5: Expression of LPAATs to Improve Sn-2 Selectivity inPrototheca moriformis

In the example we disclose genetically engineered Prototheca moriformisstrains in which we have modified fatty acid and triacylglycerolbiosynthesis to maximize the accumulation of Stearoyl-Oleoyl-Stearoyl(SOS) TAGs, and minimize the production of trisaturated TAGs. Oils fromthese strains resemble plant seed oils known as “structuring fats”,which have high proportions of Saturated-Oleate-Saturated TAGs and lowlevels of trisaturates. These structuring fats (often called “butters”)are generally solid at room temperature but melt sharply between 35-40°C.

Strains with high SOS and low trisaturates were obtained by threesuccessive transformations, beginning with S5100, a classically improvedderivative of S376 (improved to increase lipid titer), a wild typeisolate of Prototheca moriformis. S5100 was transformed with a constructto which increased expression of PmKASII-1 and ablated the SAD2-1allele. The resultant strain, S5780, produced oil with increased C18:0and lower C16:0 content relative to S5100. S5780 was prepared accordingto the methods disclosed in co-owned application WO2013/158938 and asdescribed below. C18:0 levels were increased further by transformationof S5780 with a construct overexpressing the C18:0-specific FATA1thioesterase gene from Garcinia mangostana (GarmFATA1), generatingstrain S6573. S6573 was disclosed in co-owned application WO2015/051319.Finally, accumulation of trisaturated TAGS was reduced by expression ofgenes encoding LPAATs from Brassica napus, Theobroma cacao, Garciniahombororiana or Garcinia indica in S6573 as described below.

Construct Used for SAD2 Knockout and PmKASII-1 Overexpression in S5100to Produce S5780

The sequence of the transforming DNA from the SAD2-1 ablation, PmKASIIover-expression construct, pSZ2624, is shown below. The construct iswritten as: pSZ2624:SAD2-1vD::PmKASII-1tp_PmKASII-1_FLAG-CvNR:CpACT-AtTHIC-CpEF1a::SAD2-1vERelevant restriction sites are indicated in lowercase, bold, and arefrom 5′-3′ PmeI, SpeI, AscI, ClaI, SacI, AvrII, EcoRV, AflII, KpnI,XbaI, MfeI, BamHI, BspQI and PmeI. Underlined sequences at the 5′ and 3′flanks of the construct represent genomic DNA from P. moriformis thatenable targeted integration of the transforming DNA via homologousrecombination at the SAD2-1 locus. The SAD2-1 5′ integration flankcontained the endogeneous SAD2-1 promoter, enabling the in situactivation of the PmKASII gene. Proceeding in the 5′ to 3′ direction,the region encoding the PmKASII plastid targeting sequence is indicatedby lowercase, underlined italics. The sequence that encodes the maturePmKASII polypeptide is indicated with lowercase italics, while a 3×FLAGepitope encoding sequence is in bold italics. The initiator ATG andterminator TGA for PmKASII-FLAG are indicated by uppercase italics. The3′ UTR of the Chlorella vulgaris nitrate reductase (CvNR) gene isindicated by small capitals. Two spacer regions are represented bylowercase text. The CpACT promoter driving the expression of the AtTHICgene (encoding 4-amino-5-hydroxymethyl-2-methylpyrimidine synthaseactivity, thereby permitting the strain to grow in the absence ofexogeneous thiamine) is indicated by lowercase, boxed text. Theinitiator ATG and terminator TGA for AtTHIC are indicated by uppercaseitalics, while the coding region is indicated with lowercase italics.The 3′ UTR of the Chlorella protothecoides EF1a (CpEF1a) gene isindicated by small capitals. The use of THIC as a selection marker wasdescribed in co-owned applications WO2011/150410 and WO2013/150411.

pSZ2624 Nucleotide sequence of the transforming  DNA SEQ ID NO: 86gtttaaac GCCGGTCACCACCCGCATGCTCGTACTACAGCGCACGCACCGCTTCGTGATCCACCGGGTGAACGTAGTCCTCGACGGAAACATCTGGTTCGGGCCTCCTGCTTGCACTCCCGCCCATGCCGACAACCTTTCTGCTGTTACCACGACCCACAATGCAACGCGACACGACCGTGTGGGACTGATCGGTTCACTGCACCTGCATGCAATTGTCACAAGCGCTTACTCCAATTGTATTCGTTTGTTTTCTGGGAGCAGTTGCTCGACCGCCCGCGTCCCGCAGGCAGCGATGACGTGTGCGTGGCCTGGGTGTTTCGTCGAAAGGCCAGCAACCCTAAATCGCAGGCGATCCGGAGATTGGGATCTGATCCGAGTTTGGACCAGATCCGCCCCGATGCGGCACGGGAACTGCATCGACTCGGCGCGGAACCCAGCTTTCGTAAATGCCAGATTGGTGTCCGATACCTGGATTTGCCATCAGCGAAACAAGACTTCAGCAGCGAGCGTATTTGGCGGGCGTGCTACCAGGGTTGCATACATTGCCCATTTCTGTCTGGACCGCTTTACTGGCGCAGAGGGTGAGTTGATGGGGTTGGCAGGCATCGAAACGCGCGTGCATGGTGTGCGTGTCTGTTTTCGGCTGCACGAATTCAATAGTCGGATGGGCGACGGTAGAATTGGGTGTGGCGCTCGCGTGCATGCCTCGCCCCGTCGGGTGTCATGACCGGGACTGGAATCCCCCCTCGCGACCATCTTGCTAACGCTCCCGACTCTCCCGACCGCGCGCAGGATAGACTCTTGTTCAACCAATCGACA actagt ATGcagaccgcccaccagcgcccccccaccgagggccactgatcggcgcccgcctgcccaccgcctcccgccgcgccgtgcgccgcgcctggtcccgcatcgcc cgcg ggcgcgccgccgccgccgccgacgccaaccccgcccgccccgagcgccgcgtggtgatcaccggccagggcgtggtgacctccctgggccagaccatcgagcagactactcctccctgctggagggcgtgtccggcatctcccagatccagaagacgacaccaccggctacaccaccaccatcgccggcgagatcaagtccctgcagctggacccctacgtgcccaagcgctgggccaagcgcgtggacgacgtgatcaagtacgtgtacatcgccggcaagcaggccctggagtccgccggcctgcccatcgaggccgccggcctggccggcgccggcctggaccccgccctgtgcggcgtgctgatcggcaccgccatggccggcatgacctccacgccgccggcgtggaggccctgacccgcggcggcgtgcgcaagatgaaccccactgcatccccactccatctccaacatgggcggcgccatgctggccatggacatcggatcatgggccccaactactccatctccaccgcctgcgccaccggcaactactgcatcctgggcgccgccgaccacatccgccgcggcgacgccaacgtgatgctggccggcggcgccgacgccgccatcatcccctccggcatcggcggcttcatcgcctgcaaggccctgtccaagcgcaacgacgagcccgagcgcgcctcccgcccctgggacgccgaccgcgacggatcgtgatgggcgagggcgccggcgtgctggtgctggaggagctggagcacgccaagcgccgcggcgccaccatcctggccgagctggtgggcggcgccgccacctccgacgcccaccacatgaccgagcccgacccccagggccgcggcgtgcgcctgtgcctggagcgcgccctggagcgcgcccgcctggcccccgagcgcgtgggctacgtgaacgcccacggcacctccacccccgccggcgacgtggccgagtaccgcgccatccgcgccgtgatcccccaggactccctgcgcatcaactccaccaagtccatgatcggccacctgctgggcggcgccggcgccgtggaggccgtggccgccatccaggccctgcgcaccggctggctgcaccccaacctgaacctggagaaccccgcccccggcgtggaccccgtggtgctggtgggcccccgcaaggagcgcgccgaggacctggacgtggtgctgtccaactccttcggcttcggcggccacaactcctgcgtg atcttccgcaagtacgacgag

TGA atcgatAGATCTCTTAAGGCAGCAGCAGCTCGGATAGTATCGACACACTCTGGACGCTGGTCGTGTGATGGACTGTTGCCGCCACACTTGCTGCCTTGACCTGTGAATATCCCTGCCGCTTTTATCAAACAGCCTCAGTGTGTTTGATCTTGTGTGTACGCGCTTTTGCGAGTTGCTAGCTGCTTGTGCTATTTGCGAATACCACCCCCAGCATCCCCTTCCCTCGTTTCATATCGCTTGCATCCCAACCGCAACTTATCTACGCTGTCCTGCTATCCCTCAGCGCTGCTCCTGCTCCTGCTCACTGCCCCTCGCACAGCCTTGGTTTGGGCTCCGCCTGTATTCTCCTGGTACTGCAACCTGTAAACCAGCACTGCAATGCTGATGCACGGGAAGTAGTGGGATGGGAACACAAATGGAAAGCTTAATTAAgagctccgcgtctcgaacagagcgcgcagaggaacgctgaaggtctcgcctctgtcgcacctcagcgcggcatacaccacaataaccacctgacgaatgcgcttggttcttcgtccattagcgaagcgtccggttcacacacgtgccacgttggcgaggtggcaggtgacaatgatcggtggagctgatggtcgaaacgttcacagcctaggtgatatccatcttaaggatctaagtaagattcgaagcgctcgaccgtgccggacggactgcagccccatgtcgtagtgaccgccaatgtaagtgggctggcgtaccctgtacgtgagtcaacgtcactgcacgcgcaccaccctctcgaccggcaggaccaggcatcgcgagatacagcgcgagccagacacggagtgccgagctatgcgcacgctccaactaggtaccagtttaggtccagcgtccgtggggggggacgggctgggagcttgggccgggaagggcaagacgatgcagtccctctggggagtcacagccgactgtglgtgttgcactgtgcggcccgcagcactcacacgcaaaatgcctggccgacaggcaggccctgtccagtgcaacatccacggtccctctcatcaggctcaccttgctcattgacataacggaatgcgtaccgctctttcagatctgtccatccagagaggggagcaggctccccaccgacgctgtcaaacttgcttcctgcccaaccgaaaacattattgtttgagggggggggggggggggcagattgcatggcgggatatctcgtgaggaacatcactgggacactgtggaacacagtgagtgcagtatgcagagcatgtatgctaggggtcagcgcaggaagggggcctttcccagtctcccatgccactgcaccgtatccacgactcaccaggaccagcttcttgatcggcttccgctcccgtggacaccagtglgtagcctctggactccaggtatgcgtgcaccgcaaaggccagccgatcgtgccgattcctgggtggaggatatgagtcagccaacttggggctcagagtgcacactggggcacgatacgaaacaacatctacaccgtgtcctccatgctgacacaccacagcttcgctccacctgaatgtgggcgcatgggcccgaatcacagccaatgtcgctgctgccataatgtgatccagaccctctccgcccagatgccgagcggatcgtgggcgctgaatagattcctgtttcgatcactgtttgggtcctttccttttcgtctcggatgcgcgtctcgaaacaggctgcgtcgggctttcggatcccttttgctccctccgtcaccatcctgcgcgcgggcaagttgcttgaccctgggctgataccagggttggagggtattaccgcgtcaggccattcccagcccggattcaattcaaagtctgggccaccaccctccgccgctctgtctgatcactccacattcgtgcatacactacgttcaagtcctgatccaggcgtgtctcgggacaaggtgtgcttgagtttgaatctcaaggacccactccagcacagctgctggttgaccccgccctcgcaatcta ga ATGgccgcgtccgtccactgcaccctgatgtccgtggtctgcaacaacaagaaccactccgcccgccccaagctgcccaactcctccctgctgcccggatcgacgtggtggtccaggccgcggccacccgatcaagaaggagacgacgaccacccgcgccacgctgacgacgacccccccacgaccaactccgagcgcgccaagcagcgcaagcacaccatcgacccctcctcccccgacaccagcccatcccctccacgaggagtgatccccaagtccacgaaggagcacaaggaggtggtgcacgaggagtccggccacgtcctgaaggtgcccaccgccgcgtgcacctgtccggcggcgagcccgccacgacaactacgacacgtccggcccccagaacgtcaacgcccacatcggcctggcgaagctgcgcaaggagtggatcgaccgccgcgagaagctgggcacgccccgctacacgcagatgtactacgcgaagcagggcatcatcacggaggagatgctgtactgcgcgacgcgcgagaagctggaccccgagacgtccgctccgaggtcgcgcggggccgcgccatcatcccctccaacaagaagcacctggagctggagcccatgatcgtgggccgcaagacctggtgaaggtgaacgcgaacatcggcaactccgccgtggcctcctccatcgaggaggaggtctacaaggtgcagtgggccaccatgtggggcgccgacaccatcatggacctgtccacgggccgccacatccacgagacgcgcgagtggatcctgcgcaactccgcggtccccgtgggcaccgtccccatctaccaggcgctggagaaggtggacggcatcgcggagaacctgaactgggaggtgaccgcgagacgctgatcgagcaggccgagcagggcgtggactacttcacgatccacgcgggcgtgctgctgcgctacatccccctgaccgccaagcgcctgacgggcatcgtgtcccgcggcggctccatccacgcgaagtggtgcctggcctaccacaaggagaacttcgcctacgagcactgggacgacatcctggacatctgcaaccagtacgacgtcgccctgtccatcggcgacggcctgcgccccggctccatctacgacgccaacgacacggcccagacgccgagctgctgacccagggcgagctgacgcgccgcgcgtgggagaaggacgtgcaggtgatgaacgagggccccggccacgtgcccatgcacaagatccccgagaacatgcagaagcagctggagtggtgcaacgaggcgcccactacaccctgggccccctgacgaccgacatcgcgcccggctacgaccacatcacctccgccatcggcgcggccaacatcggcgccctgggcaccgccctgctgtgctacgtgacgcccaaggagcacctgggcctgcccaaccgcgacgacgtgaaggcgggcgtcatcgcctacaagatcgccgcccacgcggccgacctggccaagcagcacccccacgcccaggcgtgggacgacgcgctgtccaaggcgcgatcgagaccgctggatggaccagacgcgctgtccctggaccccatgacggcgatgtccaccacgacgagacgctgcccgcggacggcgcgaaggtcgcccacactgctccatgtgcggccccaagactgctccatgaagatcacggaggacatccgcaagtacgccgaggagaacggctacggctccgccgaggaggccatccgccagggcatggacgccatgtccgaggagacaacatcgccaagaagacgatctccggcgagcagcacggcgaggtcggcggcgagatctacctgcccgagtcctacgtcaaggccgcgcagaag TGAcaattgACGGAGCGTCGTGCGGGAGGGAGTGTGCCGAGCGGGGAGTCCCGGTCTGTGCGAGGCCCGGCAGCTGACGCTGGCGAGCCGTACGCCCCGAGGGTCCCCCTCCCCTGCACCCTCTTCCCCTTCCCTCTGACGGCCGCGCCTGTTCTTGCATGTTCAGCGACggatcc TAGGGAGCGACGAGTGTGCGTGCGGGGCTGGCGGGAGTGGGACGCCCTCCTCGCTCCTCTCTGTTCTGAACGGAACAATCGGCCACCCCGCGCTACGCGCCACGCATCGAGCAACGAAGAAAACCCCCCGATGATAGGTTGCGGTGGCTGCCGGGATATAGATCCGGCCGCACATCAAAGGGCCCCTCCGCCAGAGAAGAAGCTCCTTTCCCAGCAGACTCCTTCTGCTGCCAAAACACTTCTCTGTCCACAGCAACACCAAAGGATGAACAGATCAACTTGCGTCTCCGCGTAGCTTCCTCGGCTAGCGTGCTTGCAACAGGTCCCTGCACTATTATCTTCCTGCTTTCCTCTGAATTATGCGGCAGGCGAGCGCTCGCTCTGGCGAGCGCTCCTTCGCGCCGCCCTCGCTGATCGAGTGTACAGTCAATGAATGGTCCTGGGCGAAGAACGAGGGAATTTGTGGGTAAAACAAGCATCGTCTCTCAGGCCCCGGCGCAGTGGCCGTTAAAGTCCAAGACCGTGACCAGGCAGCGCAGCGCGTCCGTGTGCGGGCCCTGCCTGGCGGCTCGGCGTGCCAGGCTCGAGAGCAGCTCCCTCAGGTCGCCTTGGACGGCCTCTGCGAGGCCGGTGAGGGCCTGCAGGAGCGCCTCGAGCGTGGCAGTGGCGGTCGTATCCGGGTCGCCGGTCACCGCCTGCGACTCGC CATCCgaagagcgtttaaac

Construct D1683 (pSZ2624), was transformed into S5100. Primarytransformants were clonally purified and grown under standard lipidproduction conditions at pH 5. Integration of pSZ2624 at the SAD2-1locus was verified by DNA blot analysis. The fatty acid profiles andlipid titers of lead strains were assayed in 50-mL shake flasks (Table8). Simultaneous ablation of SAD2-1 and over-expression of PmKASII(driven in situ by the SAD2-1 promoter) resulted in C18:0 levels up to26.1%. C16:0 accumulation was reduced from 15.3% in S5100 to <6% thestrains derived from D1683, demonstrating that PmKASII-1 over-expressionpromoted the elongation of C16:0 to C18:0. S5780 was chosen for furtherdevelopment as it had the highest lipid titer relative to the S5100parent.

TABLE 8 Fatty acid profiles of SAD2-1 ablation, PmKASII-1 overexpressionstrains derived from D1683-1, compared to the S5100 parent. PrimaryS5100; T531; D1683.1 Strain S5100 S5780 S5781 S5782 S5783 S5784 FattyAcid C14:0 0.7 0.7 0.8 0.7 0.7 0.7 Area % C16:0 15.3 5.9 6.0 6.0 5.8 5.8C16:1 0.5 0.1 0.0 0.1 0.0 0.0 C18:0 4.0 25.6 26.1 26.0 25.0 25.3 C18:171.0 55.7 54.5 54.6 56.3 55.6 C18:2 7.3 8.0 8.5 8.5 8.1 8.4 C18:3 α 0.50.7 0.8 0.8 0.7 0.7 C20:0 0.3 1.8 1.9 1.8 1.8 1.8 C20:1 0.2 0.6 0.6 0.60.7 0.7 C22:0 0.1 0.2 0.3 0.3 0.3 0.2 C24:0 0.1 0.4 0.4 0.4 0.4 0.4saturates 20.6 34.7 35.6 35.4 34.1 34.5

We disclose additional methods of elevating C18:0 levels that can beused in conjunction with SAD2 knockout and KASII over-expression.Previously we described acyl-ACP thioesterases from Brassica napus(BnFATA) (Co-owned application WO2012/106560), Garcinia mangostana(GarmFATA1) (Co-owned application WO2015/051319) and Theobroma cacao(TcFATA) (Co-owned application WO2013/158938) with specificity towardscleavage of C18:0-ACP, and we observed that average C18:0 levels werehigher in strains in which we replaced the native BnFATA transit peptidewith the Chlorella protothecoides SAD1 transit peptide (CpSAD1tp). A DNAconstruct was made for expression of a chimeric gene encoding CpSAD1tpfused to the predicted GarmFATA1 mature polypeptide and a FLAG tagsequence.

The sequence of the transforming DNA from the GarmFATA1 expressionconstruct pSZ3204 is shown below. The construct is written as pSZ3204:6SA::CrTUB2-ScSUC2-CvNR:PmSAD2-2-CpSAD1tp_GarmFATA1_FLAG-CvNR::6SB.Relevant restriction sites are indicated in lowercase, bold, and arefrom 5′-3′ BspQI, KpnI, XbaI, MfeI, BamHI, AvrII, EcoRV, SpeI, AscI,ClaI, AflII, SacI and BspQI. Underlined sequences at the 5′ and 3′flanks of the construct represent genomic DNA from P. moriformis thatenable targeted integration of the transforming DNA via homologousrecombination at the 6S locus. Proceeding in the 5′ to 3′ direction, theCrTUB2 promoter driving the expression of Saccharomyces cerevisiae SUC2(ScSUC2) gene, enabling strains to utilize exogeneous sucrose, isindicated by lowercase, boxed text. The initiator ATG and terminator TGAof ScSUC2 are indicated by uppercase italics, while the coding region isrepresented by lowercase italics. The 3′ UTR of the CvNR gene isindicated by small capitals. A spacer region is represented by lowercasetext. The P. moriformis SAD2-2 (PmSAD2-2) promoter driving theexpression of the chimeric CpSAD1tp_GarmFATA1_FLAG gene is indicated bylowercase, boxed text. The initiator ATG and terminator TGA areindicated by uppercase italics; the sequence encoding CpSAD1tp isrepresented by lowercase, underlined italics; the sequence encoding theGarmFATA1 mature polypeptide is indicated by lowercase italics; and the3×FLAG epitope tag is represented by uppercase, bold italics. A secondCvNR 3′ UTR is indicated by small capitals.

pSZ3204 SEQ ID NO: 87 gctcttc GCCGCCGCCACTCCTGCTCGAGCGCGCCCGCGCGTGCGCCGCCAGCGCCTTGGCCTTTTCGCCGCGCTCGTGCGCGTCGCTGATGTCCATCACCAGGTCCATGAGGTCTGCCTTGCGCCGGCTGAGCCACTGCTTCGTCCGGGCGGCCAAGAGGAGCATGAGGGAGGACTCCTGGTCCAGGGTCCTGACGTGGTCGCGGCTCTGGGAGCGGGCCAGCATCATCTGGCTCTGCCGCACCGAGGCCGCCTCCAACTGGTCCTCCAGCAGCCGCAGTCGCCGCCGACCCTGGCAGAGGAAGACAGGTGAGGGGGGTATGAATTGTACAGAACAACCACGAGCCTTGTCTAGGCAGAATCCCTACCAGTCATGGCTTTACCTGGATGACGGCCTGCGAACAGCTGTCCAGCGACCCTCGCTGCCGCCGCTTCTCCCGCACGCTTCTTTCCAGCACCGTGATGGCGCGAGCCAGCGCCGCACGCTGGCGCTGCGCTTCGCCGATCTGAGGACAGTCGGGGAACTCTGATCAGTCTAAACCCCCTTGCGCGTTAGTGTTGCCATCCTTTGCAGACCGGTGAGAGCCGACTTGTTGTGCGCCACCCCCCACACCACCTCCTCCCAGACCAATTCTGTCACCTTTTTGGCGAAGGCATCGGCCTCGGCCTGCAGAGAGGACAGCAGTGCCCAGCCGCTGGGGGTTGGCGGATGCACGCTCA ggtaccattcttgcgctatgacacttccagcaaaaggtagggcgggctgcgagacggcttcccggcgctgcatgcaacaccgatgatgcttcgaccccccgaagctccttcggggctgcatgggcgctccgatgccgctccagggcgagcgctgtttaaatagccaggcccccgattgcaaagacattatagcgagctaccaaagccatattcaaacacctagatcactaccacttctacacaggccactcgagcttgtgatcgcactccgctaagggggcgcctcttcctcttcgtttcagtcacaacccgcaaactctagaatatcaATGctgctgcaggccttcctgttcctgctggccggcttcgccgccaagatcagcgcctccatgacgaacgagacgtccgaccgccccctggtgcacttcacccccaacaagggctggatgaacgaccccaacggcctgtggtacgacgagaaggacgccaagtggcacctgtacttccagtacaacccgaacgacaccgtctgggggacgcccttgttctggggccacgccacgtccgacgacctgaccaactgggaggaccagcccatcgccatcgccccgaagcgcaacgactccggcgccttctccggctccatggtggtggactacaacaacacctccggcttcttcaacgacaccatcgacccgcgccagcgctgcgtggccatctggacctacaacaccccggagtccgaggagcagtacatctcctacagcctggacggcggctacaccttcaccgagtaccagaagaaccccgtgctggccgccaactccacccagttccgcgacccgaaggtcttctggtacgagccctcccagaagtggatcatgaccgcggccaagtcccaggactacaagatcgagatctactcctccgacgacctgaagtcctggaagctggagtccgcgttcgccaacgagggcttcctcggctaccagtacgagtgccccggcctgatcgaggtccccaccgagcaggaccccagcaagtcctactgggtgatgttcatctccatcaaccccggcgccccggccggcggctccttcaaccagtacttcgtcggcagcttcaacggcacccacttcgaggccttcgacaaccagtcccgcgtggtggacttcggcaaggactactacgccctgcagaccttcttcaacaccgacccgacctacgggagcgccctgggcatcgcgtgggcctccaactgggagtactccgccttcgtgcccaccaacccctggcgctcctccatgtccctcgtgcgcaagttctccctcaacaccgagtaccaggccaacccggagacggagctgatcaacctgaaggccgagccgatcctgaacatcagcaacgccggcccctggagccggttcgccaccaacaccacgttgacgaaggccaacagctacaacgtcgacctgtccaacagcaccggcaccctggagttcgagctggtgtacgccgtcaacaccacccagacgatctccaagtccgtgttcgcggacctctccctctggttcaagggcctggaggaccccgaggagtacctccgcatgggcttcgaggtgtccgcgtcctccttcttcctggaccgcgggaacagcaaggtgaagttcgtgaaggagaacccctacttcaccaaccgcatgagcgtgaacaaccagcccttcaagagcgagaacgacctgtcctactacaaggtgtacggcttgctggaccagaacatcctggagctgtacttcaacgacggcgacgtcgtgtccaccaacacctacttcatgaccaccgggaacgccctgggctccgtgaacatgacgacgggggtggacaacctgttctacatcgacaagttccaggtgcgcgaggtcaagT GAcaattgGCAGCAGCAGCTCGGATAGTATCGACACACTCTGGACGCTGGTCGTGTGATGGACTGTTGCCGCCACACTTGCTGCCTTGACCTGTGAATATCCCTGCCGCTTTTATCAAACAGCCTCAGTGTGTTTGATCTTGTGTGTACGCGCTTTTGCGAGTTGCTAGCTGCTTGTGCTATTTGCGAATACCACCCCCAGCATCCCCTTCCCTCGTTTCATATCGCTTGCATCCCAACCGCAACTTATCTACGCTGTCCTGCTATCCCTCAGCGCTGCTCCTGCTCCTGCTCACTGCCCCTCGCACAGCCTTGGTTTGGGCTCCGCCTGTATTCTCCTGGTACTGCAACCTGTAAACCAGCACTGCAATGCTGATGCACGGGAAGTAGTGGGATGGGAACACAAATGGAggatcccgcgtctcgaacagagcgcgcagaggaacgctgaaggtctcgcctctgtcgcacctcagcgcggcatacaccacaataaccacctgacgaatgcgcttggttcttcgtccattagcgaagcgtccggttcacacacgtgccacgttggcgaggtggcaggtgacaatgatcggtggagctgatggtcgaaacgttcacagcctagggatatcctgaagaatgggaggcaggtgttgttgattatgagtgtgtaaaagaaaggggtagagagccgtcctcagatccgactactatgcaggtagccgctcgcccatgcccgcctggctgaatattgatgcatgcccatcaaggcaggcaggcatactgtgcacgcaccaagcccacaatcttccacaacacacagcatgtaccaacgcacgcgtaaaagttggggtgctgccagtgcgtcatgccaggcatgatgtgctcctgcacatccgccatgatctcctccatcgtctcgggtgtttccggcgcctggtccgggagccgttccgccagatacccagacgccacctccgacctcacggggtacttttcgagcgtctgccggtagtcgacgatcgcgtccaccatggagtagccgaggcgccggaactggcgtgacggagggaggagagggaggagagagaggggggggggggggggggatgattacacgccagtctcacaacgcatgcaagacccgtttgattatgagtacaatcatgcactactagatggatgagcgccaggcataaggcacaccgacgttgatggcatgagcaactcccgcatcatatttcctattgtcctcacgccaagccggtcaccatccgcatgctcatattacagcgcacgcaccgcttcgtgatccaccgggtgaacgtagtcctcgacggaaacatctggctcgggcctcgtgctggcactccctcccatgccgacaacctttctgctgtcaccacgacccacgatgcaacgcgacacgacccggtgggactgatcggttcactgcacctgcatgcaattgtcacaagcgcatactccaatcgtatccgtttgatttctgtgaaaactcgctcgaccgcccgcgtcccgcaggcagcgatgacgtgtgcgtgacctgggtgtttcgtcgaaaggccagcaaccccaaatcgcaggcgatccggagattgggatctgatccgagcttggaccagatcccccacgatgcggcacgggaactgcatcgactcggcgcggaacccagcMcgtaaatgccagattggtgtccgataccttgatttgccatcagcgaaacaagacttcagcagcgagcgtatttggcgggcgtgctaccagggttgcatacattgcccatttctgtctggaccgctttaccggcgcagagggtgagttgatggggttggcaggcatcgaaacgcgcgtgcatggtgtgtgtgtctgttttcggctgcacaatttcaatagtcggatgggcgacggtagaattgggtgttgcgctcgcgtgcatgcctcgccccgtcgggtgtcatgaccgggactggaatcccccctcgcgaccctcctgctaacgctcccgactctcccgcccgcgcgcaggatagactctagttca accaatcgacaactagt ATGgccaccgcatccactactcggcgacaatgcccgctgcggcgacctgcgtcgctcggcgggctccgggccccggcgcccagcgaggcccctccccgtgcgcg ggcgcgcc atccccccccgcatcatcgtggtgtcctcctcctcctccaaggtgaaccccctgaagaccgaggccgtggtgtcctccggcctggccgaccgcctgcgcctgggctccctgaccgaggacggcctgtcctacaaggagaagacatcgtgcgctgctacgaggtgggcatcaacaagaccgccaccgtggagaccatcgccaacctgctgcaggaggtgggctgcaaccacgcccagtccgtgggctactccaccggcggataccaccacccccaccatgcgcaagctgcgcctgatctgggtgaccgcccgcatgcacatcgagatctacaagtaccccgcctggtccgacgtggtggagatcgagtcctggggccagggcgagggcaagatcggcacccgccgcgactggatcctgcgcgactacgccaccggccaggtgatcggccgcgccacctccaagtgggtgatgatgaaccaggacacccgccgcctgcagaaggtggacgtggacgtgcgcgacgagtacctggtgcactgcccccgcgagctgcgcctggccaccccgaggagaacaactcctccctgaagaagatctccaagctggaggacccctcccagtactccaagctgggcctggtgccccgccgcgccgacctggacatgaaccagcacgtgaacaacgtgacctacatcggctgggtgctggagtccatgccccaggagatcatcgacacccacgagctgcagaccatcaccctggactaccgccgcgagtgccagcacgacgacgtggtggactccctgacctcccccgagccctccgaggacgccgaggccgtgacaaccacaacggcaccaacggctccgccaacgtgtccgccaacgaccacggctgccgcaacacctgcacctgctgcgcctgtccggcaacggcctggagatcaaccgcggccgcaccgagtggcgcaagaagcccacccgc

TGA atcgatagatctcttaagGCAGCAGCAGCTCGGATAGTATCGACACACTCTGGACGCTGGTCGTGTGATGGACTGTTGCCGCCACACTTGCTGCCTTGACCTGTGAATATCCCTGCCGCTTTTATCAAACAGCCTCAGTGTGTTTGATCTTGTGTGTACGCGCTTTTGCGAGTTGCTAGCTGCTTGTGCTATTTGCGAATACCACCCCCAGCATCCCCTTCCCTCGTTTCATATCGCTTGCATCCCAACCGCAACTTATCTACGCTGTCCTGCTATCCCTCAGCGCTGCTCCTGCTCCTGCTCACTGCCCCTCGCACAGCCTTGGTTTGGGCTCCGCCTGTATTCTCCTGGTACTGCAACCTGTAAACCAGCACTGCAATGCTGATGCACGGGAAGTAGTGGGATGGGAACACAAATGGAaagcttaa ttaagagctcTTGTTTTCCAGAAGGAGTTGCTCCTTGAGCCTTTCATTCTCAGCCTCGATAACCTCCAAAGCCGCTCTAATTGTGGAGGGGGTTCGAATTTAAAAGCTTGGAATGTTGGTTCGTGCGTCTGGAACAAGCCCAGACTTGTTGCTCACTGGGAAAAGGACCATCAGCTCCAAAAAACTTGCCGCTCAACACCGCGTACCTCTGCTTTGCGCAATCTGCCCTGTTGAAATCGCCACCACATTCATATTGTGACGCTTGAGCAGTCTGTAATTGCCTCAGAATGTGGAATCATCTGCCCCCTGTGCGAGCCCATGCCAGGCATGTCGCGGGCGAGGACACCCGCCACTCGTACAGCAGACCATTATGCTACCTCACAATAGTTCATAACAGTGACCATATTTCTCGAAGCTCCCCAACGAGCACCTCCATGCTCTGAGTGGCCACCCCCCGGCCCTGGTGCTTGCGGAGGGCAGGTCAACCGGCATGGGGCTACCGAAATCCCCGACCGGATCCCACCACCCCCGCGATGGGAAGAATCTCTCCCCGGGATGTGGGCCCACCACCAGCACAACCTGCTGGCCCAGGCGAGCGTCAAACCATACCACACAAATATCCTTGGCATCGGCCCTGAATTCCTTCTGCCGCTCTGCTACCCGGTGCTTCTGTCCGAAGCAGGGGTTGCTAGGGATCGCTCCGAGTCCGCAAACCCTTGTCGCGTGGCGGGGCTT GTTCGAGCTT gaagagc

Construct D1940 (pSZ3204), was transformed into the S5780 parent strain.Primary transformants were clonally purified and grown under standardlipid production conditions at pH 5. Integration of pSZ3204 at the 6Slocus was verified by DNA blot analysis. The fatty acid profiles andlipid titers of lead strains were assayed in 50-mL shake flasks (Table9). Over-expression of GarmFATA1 (driven by the SAD2-2 promoter)resulted in C18:0 levels up to 54.3%. C16:0 levels were comparable instrains derived from D1940 and the S5780 parent. S6573 was chosen forfurther development as it had the highest lipid titer of the strainswith >50% C18:0.

TABLE 9 Fatty acid profiles of GarmFATA1 overexpressing stable strainsderived from D1940 primary transformants. Primary D1683.1 D1940.19D1940.20 D1940.23 D1940.46 D1940.5 Strain S5100 S5780 S6571 S6572 S6573S6574 S6575 S6578 S6580 Fatty Acid Area % C14:0 0.7 0.0 0.8 0.0 0.8 0.70.7 0.0 0.0 C16:0 18.0 5.9 6.3 6.6 6.3 5.0 5.1 5.0 5.3 C16:1 0.5 0.0 0.10.1 0.1 0.0 0.1 0.1 0.1 C18:0 3.9 29.0 52.7 54.3 53.7 43.1 46.0 45.447.9 C18:1 69.8 54.3 31.4 30.1 30.5 41.5 38.5 40.0 37.2 C18:2 5.9 6.45.7 5.8 5.6 6.3 6.2 6.1 6.2 C18:3 α 0.5 0.7 0.6 0.6 0.6 0.6 0.5 0.6 0.5C20:0 0.3 2.4 1.8 1.6 1.7 2.1 2.0 2.0 2.0 C20:1 0.1 0.6 0.1 0.1 0.1 0.20.1 0.1 0.1 C22:0 0.1 0.3 0.2 0.2 0.2 0.3 0.3 0.2 0.2 C24:0 0.0 0.0 0.00.0 0.0 0.0 0.0 0.0 0.0 saturates 23.1 37.7 61.9 62.8 62.8 51.2 54.252.7 55.5

Lysophosphatidic acid acetyltransferase (LPAAT) enzymes are responsiblefor the transfer of acyl groups to the sn-2 position on the glycerolbackbone. We disclose here that we can reduce the accumulation ofexcessive amounts of trisaturates in our high SOS strains by expressingheterologous LPAAT genes which were better than the endogenousacyltransferases at discriminating against saturated fatty acids.Expression of LPAT2 homologs from B. napus, T cacao, Garciniahombroriana and Garcinia indica and their effect on the formation oftrisaturated TAGs in the high-C18:0 S6573 strain is disclosed below.

The sequence of the transforming DNA from the BnLPAT2(Bn1.13) expressionconstruct pSZ4198 is shown below The construct is written as pSZ4198:PLOOP::PmHXT1-ScarMEL1-CvNR:PmSAD2-2v2-BnLPAT2(Bn1.13)-CvNR::PLOOP.Relevant restriction sites are indicated in lowercase, bold, and arefrom 5′-3′ BspQI, KpnI, SpeI, SnaBI, EcoRI, SpeI, ClaI, BglII, AflII,HindIII, SacI and BspQI. Underlined sequences at the 5′ and 3′ flanks ofthe construct represent genomic DNA from P. moriformis that enabletargeted integration of the transforming DNA via homologousrecombination at the PLOOP locus. Proceeding in the 5′ to 3′ direction,the PmHXT1 promoter driving the expression of S. carlbergensis MEL1(ScarMEL1) gene, enabling strains to utilize exogeneous melibiose, isindicated by lowercase, boxed text. The initiator ATG and terminator TGAof ScarMEL1 are indicated by uppercase italics, while the coding regionis represented by lowercase italics. The 3′ UTR of the CvNR gene isindicated by small capitals. The P. moriformis SAD2-2v2 promoter drivingthe expression of the BnLPAT2(Bn1.13) gene is indicated by lowercase,boxed text. The initiator ATG and terminator TGA are indicated byuppercase italics; the sequence encoding BnLPAT2(Bn1.13) is representedby lowercase, underlined italics. A second CvNR 3′ UTR is indicated bysmall capitals. The Brassica napus LPAAT2(BN1.13) sequence is fromGenbank accession GU045434.

SEQ ID NO: 88: Nucleotide sequence of the transforming DNA from pSZ4198gctcttccgctAACGGAGGTCTGTCACCAAATGGACCCCGTCTATTGCGGGAAACCACGGCGATGGCACGTTTCAAAACTTGATGAAATACAATATTCAGTATGTCGCGGGCGGCGACGGCGGGGAGCTGATGTCGCGCTGGGTATTGCTTAATCGCCAGCTTCGCCCCCGTCTTGGCGCGAGGCGTGAACAAGCCGACCGATGTGCACGAGCAAATCCTGACACTAGAAGGGCTGACTCGCCCGGCACGGCTGAATTACACAGGCTTGCAAAAATACCAGAATTTGCACGCACCGTATTCGCGGTATTTTGTTGGACAGTGAATAGCGATGCGGCAATGGCTTGTGGCGTTAGAAGGTGCGACGAAGGTGGTGCCACCACTGTGCCAGCCAGTCCTGGCGGCTCCCAGGGCCCCGATCAAGAGCCAGGACATCCAAACTACCCACAGCATCAACGCCCCGGCCTATACTCGAACCCCACTTGCACTCTGCAATGGTATGGGAACCACGGGGCAGTCTTGTGTGGGTCGCGCCTATCGCGGTCGGCGAA GACCGGGAAggtaccgcggtgagaatcgaaaatgcatcgtttctaggttcggagacggtcaattccctgctccggcgaatctgtcggtcaagctggccagtggacaatgttgctatggcagcccgcgcacatgggcctcccgacgcggccatcaggagcccaaacagcgtgtcagggtatgtgaaactcaagaggtccctgctgggcactccggccccactccgggggcgggacgccaggcattcgcggtcggtcccgcgcgacgagcgaaatgatgattcggttacgagaccaggacgtcgtcgaggtcgagaggcagcctcggacacgtctcgctagggcaacgccccgagtccccgcgagggccgtaaacattgtttctgggtgtcggagtgggcattttgggcccgatccaatcgcctcatgccgctctcgtctggtcctcacgttcgcgtacggcctggatcccggaaagggcggatgcacgtggtgttgccccgccattggcgcccacgtttcaaagtccccggccagaaatgcacaggaccggcccggctcgcacaggccatgctgaacgcccagatttcgacagcaacaccatctagaataatcgcaaccatccgcgttttgaacgaaacgaaacggcgctgtttagcatgtttccgacatcgtgggggccgaagcatgctccggggggaggaaagcgtggcacagcggtagcccattctgtgccacacgccgacgaggaccaatccccggcatcagccttcatcgacggctgcgccgcacatataaagccggacgcctaaccggtttcgtggttatgactagtATGttcgcgttctacttcctgacggcctgcatctccctgaagggcgtgacggcgtctccccctcctacaacggcctgggcctgacgccccagatgggctgggacaactggaacacgacgcctgcgacgtctccgagcagctgctgctggacacggccgaccgcatctccgacctgggcctgaaggacatgggctacaagtacatcatcctggacgactgctggtcctccggccgcgactccgacggcacctggtcgccgacgagcagaagaccccaacggcatgggccacgtcgccgaccacctgcacaacaactccacctgacggcatgtactcctccgcgggcgagtacacgtgcgccggctaccccggctccctgggccgcgaggaggaggacgcccagacttcgcgaacaaccgcgtggactacctgaagtacgacaactgctacaacaagggccagacggcacgcccgagatctcctaccaccgctacaaggccatgtccgacgccctgaacaagacgggccgccccatcttctactccctgtgcaactggggccaggacctgaccttctactggggctccggcatcgcgaactcctggcgcatgtccggcgacgtcacggcggagacacgcgccccgactcccgctgcccctgcgacggcgacgagtacgactgcaagtacgccggcaccactgctccatcatgaacatcctgaacaaggccgcccccatgggccagaacgcgggcgtcggcggctggaacgacctggacaacctggaggtcggcgtcggcaacctgacggacgacgaggagaaggcgcacttaccatgtgggccatggtgaagtcccccctgatcatcggcgcgaacgtgaacaacctgaaggcctcctcctactccatctactcccaggcgtccgtcatcgccatcaaccaggactccaacggcatccccgccacgcgcgtctggcgctactacgtgtccgacacggacgagtacggccagggcgagatccagatgtggtccggccccctggacaacggcgaccaggtcgtggcgctgctgaacggcggctccgtgtcccgccccatgaacacgaccctggaggagatcacttcgactccaacctgggctccaagaagctgacctccacctgggacatctacgacctgtgggcgaaccgcgtcgacaactccacggcgtccgccatcctgggccgcaacaagaccgccaccggcatcctgtacaacgccaccgagcagtcctacaaggacggcctgtccaagaacgacacccgcctgacggccagaagatcggctccctgtcccccaacgcgatcctgaacacgaccgtccccgcccacggcatcgcgactaccgcctgcgcccctcctccTGAtacgtactcgagGCAGCAGCAGCTCGGATAGTATCGACACACTCTGGACGCTGGTCGTGTGATGGACTGTTGCCGCCACACTTGCTGCCTTGACCTGTGAATATCCCTGCCGCTTTTATCAAACAGCCTCAGTGTGTTTGATCTTGTGTGTACGCGCTTTTGCGAGTTGCTAGCTGCTTGTGCTATTTGCGAATACCACCCCCAGCATCCCCTTCCCTCGTTTCATATCGCTTGCATCCCAACCGCAACTTATCTACGCTGTCCTGCTATCCCTCAGCGCTGCTCCTGCTCCTGCTCACTGCCCCTCGCACAGCCTTGGTTTGGGCTCCGCCTGTATTCTCCTGGTACTGCAACCTGTAAACCAGCACTGCAATGCTGATGCACGGGAAGTAGTGGGATGGGAACACAAATGGAAagctgtagaattcctggctcgggcctcgtgctggcactccctcccatgccgacaacctttctgctgtcaccacgacccacgatgcaacgcgacacgacccggtgggactgatcggttcactgcacctgcatgcaattgtcacaagcgcatactccaatcgtatccgtttgatttctgtgaaaactcgctcgaccgcccgcgtcccgcaggcagcgatgacgtgtgcgtgacctgggtgtttcgtcgaaaggccagcaaccccaaatcgcaggcgatccggagattgggatctgatccgagcttggaccagatcccccacgatgcggcacgggaactgcatcgactcggcgcggaacccagctttcgtaaatgccagattggtgtccgataccttgatttgccatcagcgaaacaagacttcagcagcgagcgtatttggcgggcgtgctaccagggttgcatacattgcccatttctgtctggaccgctttaccggcgcagagggtgagttgatggggttggcaggcatcgaaacgcgcgtgcatggtgtgtgtgtctgttttcggctgcacaatttcaatagtcggatgggcgacggtagaattgggtgttgcgctcgcgtgcatgcctcgccccgtcgggtgtcatgaccgggactggaatcccccctcgcgaccctcctgctaacgctcccgactctcccgcccgcgcgcaggatagactctagttcaaccaatcgacaactagtATGgccatggccgccgccgtgatcgtgcccctgggcatcctgacttcatctccggcctggtggtgaacctgctgcaggccatctgctacgtgctgatccgccccctgtccaagaacacctaccgcaagatcaaccgcgtggtggccgagaccctgtggctggagctggtgtggatcgtggactggtgggccggcgtgaagatccaggtgacgccgacaacgagaccacaaccgcatgggcaaggagcacgccctggtggtgtgcaaccaccgctccgacatcgactggctggtgggctggatcctggcccagcgctccggctgcctgggctccgccctggccgtgatgaagaagtcctccaagacctgcccgtgatcggctggtccatgtggactccgagtacctgacctggagcgcaactgggccaaggacgagtccaccctgaagtccggcctgcagcgcctgaacgacttcccccgcccataggctggccctgacgtggagggcacccgatcaccgaggccaagctgaaggccgcccaggagtacgccgcctcctccgagctgcccgtgccccgcaacgtgctgatcccccgcaccaagggcttcgtgtccgccgtgtccaacatgcgctccttcgtgcccgccatctacgacatgaccgtggccatccccaagacctcccccccccccaccatgctgcgcctgacaagggccagccctccgtggtgcacgtgcacatcaagtgccactccatgaaggacctgcccgagtccgacgacgccatcgcccagtggtgccgcgaccagacgtggccaaggacgccctgctggacaagcacatcgccgccgacaccaccccggccagcaggagcagaacatcggccgccccatcaagtccctggccgtggtgctgtcctggtcctgcctgctgatcctgggcgccatgaagacctgcactggtccaacctgactcctcctggaagggcatcgccactccgccctgggcctgggcatcatcaccctgtgcatgcagatcctgatccgctcctcccagtccgagcgctccacccccgccaaggtggtgcccgccaagcccaaggacaaccacaacgactccggctcctcctcccagaccgaggtggagaagcagaagTGA atcgatagatctcttaagGCAGCAGCAGCTCGGATAGTATCGACACACTCTGGACGCTGGTCGTGTGATGGACTGTTGCCGCCACACTTGCTGCCTTGACCTGTGAATATCCCTGCCGCTTTTATCAAACAGCCTCAGTGTGTTTGATCTTGTGTGTACGCGCTTTTGCGAGTTGCTAGCTGCTTGTGCTATTTGCGAATACCACCCCCAGCATCCCCTTCCCTCGTTTCATATCGCTTGCATCCCAACCGCAACTTATCTACGCTGTCCTGCTATCCCTCAGCGCTGCTCCTGCTCCTGCTCACTGCCCCTCGCACAGCCTTGGTTTGGGCTCCGCCTGTATTCTCCTGGTACTGCAACCTGTAAACCAGCACTGCAATGCTGATGCACGGGAAGTAGTGGGATGGGAACACAAATGGAaagcttaattaagagctc AGCGGCGACGGTCCTGCTACCGTACGACGTTGGGCACGCCCATGAAAGTTTGTATACCGAGCTTGTTGAGCGAACTGCAAGCGCGGCTCAAGGATACTTGAACTCCTGGATTGATATCGGTCCAATAATGGATGGAAAATCCGAACCTCGTGCAAGAACTGAGCAAACCTCGTTACATGGATGCACAGTCGCCAGTCCAATGAACATTGAAGTGAGCGAACTGTTCGCTTCGGTGGCAGTACTACTCAAAGAATGAGCTGCTGTTAAAAATGCACTCTCGTTCTCTCAAGTGAGTGGCAGATGAGTGCTCACGCCTTGCACTTCGCTGCCCGTGTCATGCCCTGCGCCCCAAAATTTGAAAAAAGGGATGAGATTATTGGGCAATGGACGACGTCGTCGCTCCGGGAGTCAGGACCGGCGGAAAATAAGAGGCAACACACTC CGCTTCTTA gctcttc

Additional transforming constructs to test the activity of LPAATs fromB. napus, T cacao, G. hombroriana and G. indica contained the sameselectable marker, restriction sites, promoters and 3′ UTR elements aspSZ4198. The coding sequences of BnLPAT2(Bn1.5), TcLPAT2, GhomLPAT2A,GhomLPAT2B, GhomLPAT2C, GindLPAT2A, GindLPAT2B and GindLPAT2C are shownin below. In each case the initiator ATG and terminator TGA areindicated by uppercase italics; the sequence encoding the LPAT2 homologis represented by lowercase italics. The Brassica napus LPAAT2(BN1.13)sequence is from Genbank accession GU045435. The Theobroma cacao LPAAT2sequence is from the cocoaGenDB database.

SEQ ID NO: 89 Nucleotide sequence of the BnLPAT2(1.5) coding sequence,used in the transforming DNA from pSZ4202ATGgccatggccgccgccgccgtgatcgtgcccctgggcatcctgacttcatctccggcctggtggtgaacctgctgcaggccgtgtgctacgtgctgatccgccccctgtccaagaacacctaccgcaagatcaaccgcgtggtggccgagaccctgtggctggagctggtgtggatcgtggactggtgggccggcgtgaagatccaggtgacgccgacgacgagaccacaaccgcatgggcaaggagcacgccctggtggtgtgcaaccaccgctccgacatcgactggctggtgggctggatcctggcccagcgctccggctgcctgggctccgccctggccgtgatgaagaagtcctccaagacctgcccgtgatcggctggtccatgtggactccgagtacctgacctggagcgcaactgggccaaggacgagtccaccctgaagtccggcctgcagcgcctgaacgacacccccgccccactggctggccctgacgtggagggcacccgatcaccgaggccaagctgaaggccgcccaggagtacgccgcctcctcccagctgcccgtgccccgcaacgtgctgatcccccgcaccaagggcttcgtgtccgccgtgtccaacatgcgctccttcgtgcccgccatctacgacatgaccgtggccatccccaagacctcccccccccccaccatgctgcgcctgacaagggccagccctccgtggtgcacgtgcacatcaagtgccactccatgaaggacctgcccgagtccgacgacgccatcgcccagtggtgccgcgaccagacgtggccaaggacgccctgctggacaagcacatcgccgccgacaccaccccggccagaaggagcacaacatcggccgccccatcaagtccctggccgtggtggtgtcctgggcctgcctgctgaccctgggcgccatgaagacctgcactggtccaacctgactcctccctgaagggcatcgccctgtccgccctgggcctgggcatcatcaccctgtgcatgcagatcctgatccgctcctcccagtccgagcgctccacccccgccaaggtggcccccgccaagcccaaggacaagcaccagtccggctcctcctcccagaccgaggtggaggagaagcagaagTGASEQ ID NO: 90 Nucleotide sequence of the TcLPAT2 coding sequence, usedin the transforming DNA from pSZ4206ATGgccatcgccgccgccgccgtgatcgtgcccctgggcctgctgttcttcatctccggcctggtggtgaacctgatccaggccctgtgcttcgtgctgatccgccccctgtccaagaacacctaccgcaagatcaaccgcgtggtggccgagctgctgtggctggagctgatctggctggtggactggtgggccggcgtgaagatcaaggtgttcatggaccccgagtccttcaacctgatgggcaaggagcacgccctggtggtggccaaccaccgctccgacatcgactggctggtgggctggctgctggcccagcgctccggctgcctgggctccgccctggccgtgatgaagaagtcctccaagttcctgcccgtgatcggctggtccatgtggttctccgagtacctgttcctggagcgctcctgggccaaggacgagaacaccctgaaggccggcctgcagcgcctgaaggacttcccccgccccttctggctggccttcttcgtggagggcacccgcttcacccaggccaagttcctggccgcccaggagtacgccgcctcccagggcctgcccatcccccgcaacgtgctgatcccccgcaccaagggcttcgtgtccgccgtgtcccacatgcgctccttcgtgcccgccatctacgacatgaccgtggccatccccaagtcctccccctcccccaccatgctgcgcctgttcaagggccagccctccgtggtgcacgtgcacatcaagcgctgcctgatgaaggagctgcccgagaccgacgaggccgtggcccagtggtgcaaggacatgttcgtggagaaggacaagctgctggacaagcacatcgccgaggacaccttctccgaccagcccatgcaggacctgggccgccccatcaagtccctgctggtggtggcctcctgggcctgcctgatggcctacggcgccctgaagttcctgcagtgctcctccctgctgtcctcctggaagggcatcgccttcttcctggtgggcctggccatcgtgaccatcctgatgcacatcctgatcctgttctcccagtccgagcgctccacccccgccaaggtggcccccggcaagcccaagaacgacggcgagacctccgaggcccgccgcgacaagcagcagTGASEQ ID NO: 91 Nucleotide sequence of the GhomLPAT2A coding sequence,used in the transforming DNA from pSZ4412.ATGgccatccccgccgccatcgtgatcgtgcccgtgggcctgctgttcttcatctccggcctgatcgtgaacctgctgcaggccctgtgcttcgtgctgatccgccccctgtccaagtccgcctaccgcaccatcaaccgccagctggtggagctgctgtggctggagctggtgtgcatcgtggactggtgggcccgcgtgaagatccagctgttcaccgacaaggagaccctgaactccatgggcaaggagcacgccctggtgatgtgcaaccaccgctccgacatcgactggctggtgggctggatcctggcccagcgctccggctgcctgggctccaccgtggccgtgatgaagaagtcctccaaggtgctgcccgtgatcggctggtccatgtggttctccgagtacctgttcctggagcgcaactgggccaaggacgagtccaccctgaagtccggcctgcagcgcctgcgcgacttcccccgccccttctggctggccctgttcgtggagggcacccgcttcacccagcccaagctgctggccgcccaggagtacgccgcctccaccggcctgcccatcccccgcaacgtgctgatcccccgcaccaagggcttcgtgtccgccgtgtccatcacccgctccttcgtgcccgtgatctacgacatcaccgtggccatccccaagtcctccccccagcccaccatgctgcgcctgttcaagggccagtcctccgtggtgcacgtgcacctgaagcgccacctgatgaaggacctgcccgagtccgacgacgacgtggcccagtggtgccgcgaccagttcgtggtgaaggactccctgctggacaagcacatcgccgaggacaccttctccgaccaggagctgcaggacatcggccgccccatcaagtccctggtggtgttcacctcctgggtgtgcatcatcaccttcggcgccctgaagttcctgcagtggtcctccctgctgcactcctggaagggcatcgccatctccgcctccggcctggccatcgtgaccgtgctgatgcacatcctgatccgcttctcccagtccgagcactccacctccgccaagatcgccgccgagaagcacaagaacggcggcgtgtcccaggagatgggccgcgagaagcagcacTGASEQ ID NO: 92 Nucleotide sequence of the GhomLPAT2B coding sequence,used in the transforming DNA from pSZ4413.ATGgagatccccgccgtggccgtgatcgtgcccatcggcatcctgttcttcatctccggcctgatcgtgaacctgatgcaggccatctgcttcttcctgatccgccccctgtccaagaacacccaccgcatcgtgaaccgccagctggccgagctgctgtggctggagctgatctggatcgtggactggtgggccggcgtgaagatccagctgttcaccgacaaggagaccctgcacctgatgggcaaggagcacgccctggtgatctgcaaccactcctccgacatcgactggctggtgggctggctgctgtgccagcgctccggctgcctgggctccgccctggccgtgatgaagtcctcctccaaggtgctgcccgtgatcggctggtccatgtggttctccgagtacctgttcctggagcgctcctgggccaaggacgagtccaccctgaagtccggcctgcagcgcctgaaggacttcccccgccccttctggctggccctgttcgtggagggcacccgcttcacccaggccaagctgctggccgcccaggagtacgccatgtccgccggcctgcccgtgccccgcaacgtgctgatcccccgcaccaagggcttcgtgtccgccgtgtccaacatgcgctccttcgtgcccgccatctacgacgtgaccgtggccatccccaagtcctccgtgcagcccaccatgctgcgcctgttcaagggccagtcctccgtggtgcaggtgcacctgaagcgccactccatgaaggacctgcccgagtccgaggacgacgtggcccagtggtgccgcgaccgcttcgtggtgaaggactccctgctggacaagcacaaggtggaggacaccttcaccgaccaggagctgcaggacctgggccgccccatcaagtccctggtggtggtgacctgctgggcctgcatcatcatcttcggcatcctgaagttcctgcagtggtcctccctgctgtactcctggaagggcatggccatctccgcctccggcctggccgtggtgaccttcctgatgcagatcctgatccgcttctcccagtccgagcgctccacccccgccaagatcgcccccgccaagcccaacaaggccggcaactcctccgagaccgtgcgcgacaagcaccagTGASEQ ID NO: 93 Nucleotide sequence of the GhomLPAT2C coding sequence,used in the transforming DNA from pSZ4414.ATGgccatccccgccgccatcatcatcgtgcccctgggcctgatcttcttcacctccggcctgatcatcaacctgatccaggccgtgtgctacgtgctgatccgccccctgtccaagtccaccttccgccgcatcaaccgcgagctggccgagctgctgtggctggagctggtgtgggtggtggactggtgggccggcgtgaagatccagctgttcaccgacaaggagaccctgcactccatgggcaaggagcacgccctggtgatctgcaaccaccgctccgacatcgactggctggtgggctggatcctggcccagcgctccggctgcctgggctccgccctggccgtgatgaagaagtcctccaaggtgctgcccgtgatcggctggtccatgtggttctccgagtacttcttcctggagcgcaactgggccatggacgagtccaccctgaagtccggcctgcagcgcctgaaggacttcccccagcccttctggctggccctgttcgtggagggcacccgcttcacccagcccaagctgctggccgcccaggagtacgccgcctccgccggcctgcccatcccccgcaacgtgctgatcccccgcaccaagggcttcgtgtccgccgtgaacatcatgcgctccttcgtgcccgccatctacgacgtgaccgtggccatccccaagtcctccccccagcccaccatgctgcgcctgttcaagggccagtcctccgtggtgcacgtgcacctgaagcgccacctgatggaggacctgcccgagaccgacgacgacgtggcccagtggtgccgcgaccgcttcgtggtgaaggactccctgctggacaagtacgtggccgaggacaccttctccgaccaggagctgcaggacctgggccgccccatcaagtccctggtggtggtgacctcctgggtgtgcatcatcgccttcggctccctgaagttcctgcagtggtcctccctgctgtactcctggaagggcatcgtgatctccgccgcctccctggccgtggtgaccgtgctgatgcagatcctgatccgcttctcccagtccgagcgctccacctccgccaagatcgccgccgccaagcgcaagaacgtgggcgagcacTGASEQ ID NO: 94 Nucleotide sequence of the GindPAT2A coding sequence,used in the transforming DNA from pSZ4415.ATGgccatccccgtggtggtggtgatcgtgcccgtgggcctgctgttcttcatctccggcctgatcgtgaacctgctgcaggccctgtgcttcgtgctgatccgccccctgtccaagtccgcctaccgcaccatcaaccgccagctggtggagctgctgtggctggagctggtgtgcatcgtggactggtgggcccgcgtgaagatccagctgttcatcgacaaggagaccctgaactccatgggcaaggagcacgccctggtgatgtgcaaccaccgctcctacatcgactggctggtgggctggatcctggcccagcgctccggctgcctgggctccaccgtggccgtgatgaagaagtcctccaaggtgctgcccgtgatcggctggtccatgtggttctccgagtacctgttcctggagcgcaactgggccaaggacgagtccaccctgaagtccggcctgcagcgcctgcgcgacttcccccgccccttctggctggccctgttcgtggagggcacccgcttcacccagcccaagctgctggccgcccaggagtacgccgcctccaccggcctgcccatcccccgcaacgtgctgatcccccgcaccaagggcttcgtgtccgccgtgtccatcacccgctccttcgtgcccgtgatctacgacatcaccgtggccatccccaagtcctcctcccagcccaccatgctgaagctgttcaagggccagtcctccgtggtgcacgtgcacctgaagcgccacctgatgaaggacctgcccgagtccgacgacgacgtggcccagtggtgccgcgcccagttcgtggtgaaggactccctgctggacaagcacatcgccgaggacaccttctccgaccaggagctgcaggacatcggccgccccatcaagtccctggtggtgttcacctcctgggtgtgcatcatcaccttcggcgccctgaagttcctgcagtggtcctccctgctgcactcctggaagggcatcgccatctccgcctccggcctggccatcgtgaccgtgctgatgcacatcctgatccgcttctcccagtccgagcactccacctccgccaagatcgccgccgagaagcacaagaacggcggcgtgtcccaggagatgggccgcgagaagcagcacTGASEQ ID NO: 95 Nucleotide sequence of the GindPAT2B coding sequence,used in the transforming DNA from pSZ4416.ATGggcatccccgccgtggccgtgatcgtgcccatcggcatcctgttcttcatctccggcttcatcgtgaacctgatgcaggccatctgcttcgtgctgatccgccccctgtccaagaacacctaccgcatcgtgaaccgccagctggccgagttcctgtggctggagctgatctgggtggtggactggtgggccggcgtgaagatccagctgttcaccgacaaggagaccctgcacctgatgggcaaggagcacgccctggtgatctgcaaccaccgctccgacatcgactggctggtgggctggctgctgtgccagcgctccggctgcctgggctccgccctggccgtgatgaagtcctcctccaaggtgctgcccgtgatcggctggtccatgtggttctccgagtacctgttcctggagcgctcctgggccaaggacgagtccaccctgaagctgggcctgcagcgcctgaaggacttcccccgccccttctggctggccctgttcgtggagggcacccgcttcacccaggccaagctgctggccgcccaggagtacgccatgtccgccggcctgcccgtgccccgcaacgtgctgatcccccgcaccaagggcttcgtgtccgccgtgtccaacatgcgctccttcgtgcccgccatctacgacgtgaccgtggccatccccaagtcctccgtgcagcccaccatgctgggcctgttcaagggccagtcctgcgtggtgcaggtgcacctgaagcgccacctgatgaaggacctgcccgagtccgaggacgacgtggcccagtggtgccgcgagcgcttcgtggtgaaggactccctgctggacaagcacaaggtggaggacaccttctccgaccaggagctgcaggacctgggccgccccatcaagtccctggtggtggtgatctcctgggcctgcatcctgatcttctggatcctgaagttcctgcagtggtcctccctgctgtactcctggaagggcatcgccatctccgcctgcgccatggccgtgatcgccttcctgatgcagatcctgctgcgcttctcccagtccgagcgctccacccccgccaagatcgcccccgccaagcccaacaacgcccgcaactcctccgagaccgtgcgcgacaagcaccagTGASEQ ID NO: 96 Nucleotide sequence of the GindPAT2C coding sequence,used in the transforming DNA from pSZ4417.ATGgccatccccgccgccatcatcatcgtgcccctgggcctgatcttcttcacctccggcttcatcatcaacctgatccaggccgtgtgctacgtgctgatccgccccctgtccaagtccaccttccgccgcatcaaccgccagctggccgagctgctgtggctggagctggtgtgggtggtggactggtgggccggcgtgaagatccagctgttcaccaacaaggagaccctgcactccatcggcaaggagcacgccctggtgatctgcaaccagcgctccgacatcgactggctggtgggctggatcctggcccagcgctccggctgcctgggctccgccctggccgtgatgaagaagtcctccaaggtgctgcccgtgatcggctggtccatgtggttctccgagtacctgttcctggagcgcaactgggccatggacgagtccaccctgaagtccggcctgcagtggctgaaggacttcccccagcccttctggctggccctgttcgtggagggcacccgcttcacccagcccaagctgctggccgcccaggagtacgccgcctccgccggcctgcccatcccccgcaacgtgctgatcccccgcaccaagggcttcgtgtccgccgtgaacatcatgcgctccttcgtgcccgccgtgtacgacgtgaccgtggccatccccaagtcctccccccagcccaccatgctgcgcctgttcaagggccagtcctccgtggtgcacgtgcacctgaagcgccacctgatggaggacctgcccgagaccgacgacgacgtggcccagtggtgccgcgaccgcttcgtggtgaaggactccctgctggacaagcacctggccgaggacaccttctccgaccaggagctgcaggacctgggccgccccatcaagtccctggtggtggtgacctcctgggtgtgcatcatcgccttcggcgccctgaagttcctgcagtggtcctccctgctgtactcctggaagggcatcgtgatctccgccgcctccctggccgtggtgaccgtgctgatgcagatcctgatccgcttctcccagtccgagcgctccacctccgccaaggtggtggccgagaagcgcaagaacgtgggcgagcacTGA

Constructs D2971, D2973, D2975, D3219, D3221, D3223, D3225, D3227 andD3229, derived from pSZ4198, pSZ4202, pSZ4206, pSZ4412, pSZ4413,pSZ4414, pSZ4415, pSZ4416 and pSZ4417, respectively, were transformedinto the S6573 parent strain. The fatty acid profiles of primarytransformants are shown in Table 10. Also shown are the SOS/SSS ratiosdetermined by LC/MS multiple response measurements. Expression of LPAT2genes had no discernable effect on C16:0 or C18:0 accumulation, butC18:2 levels increased by 1-2% compared to the S6573 parent in strainswhen expressing the D2971, D2973, D2975, D3221, D3223, and D3227constructs. Expression of LPAT2 genes increased C18:2 and also elevatedratios of SOS/SSS, showing reduced accumulation of trisaturated TAGs.

TABLE 10 Fatty acid profiles and SOS/SSS ratios of D2971, D2973, D2975,D3219, D3221, D3223, D3225, D3227 and D3229 primary transformants.Strain LPAAT gene SOS/SSS C14:0 C16:0 C18:0 C18:1 C18:2 C18:3 α C20:0saturates S5100 0.7 17.7 4.1 68.5 6.8 0.6 0.4 23.3 S6573.1 15 0.8 6.250.7 33.7 5.6 0.7 1.5 59.8 D2971.1 BnLPAT2(1.13) 23 0.8 6.1 51.4 30.58.6 0.6 1.4 60.2 D2971.2 16 0.8 6.1 54.3 28.9 7.0 0.6 1.5 63.3 D2971.416 0.8 6.4 53.3 29.5 7.3 0.6 1.4 62.6 S6573.2 14 0.8 6.6 52.8 31.7 5.20.6 1.5 62.3 D2973.2 BnLPAT2(1.5) 22 0.8 6.2 53.4 28.3 6.4 0.6 1.7 62.7D2973.38 23 0.9 7.5 51.2 29.1 6.5 0.5 1.4 61.7 D2973.24 24 0.9 6.8 51.729.2 6.3 0.5 1.6 61.5 S6573.3 14 0.8 6.6 52.8 31.7 5.2 0.6 1.5 62.3D2975.33 TcLPAT2 27 0.8 6.6 52.7 29.7 7.1 0.6 1.5 62.3 D2975.13 32 0.86.5 52.4 30.2 7.3 0.6 1.4 61.7 D2975.35 27 0.8 6.5 52.8 29.6 7.3 0.6 1.562.2 S6573.4 12 0.9 6.4 54.9 28.9 5.7 0.6 1.7 64.5 D3219.19 GhomLPAT2A12 0.9 7.1 52.4 31.2 4.8 0.5 2.0 63.1 D3219.20 14 0.9 6.6 53.2 30.6 5.50.6 1.7 63.0 D3219.32 15 0.8 6.4 53.1 29.8 6.5 0.6 1.5 62.6 S6573.5 120.9 6.4 53.7 30.3 5.5 0.6 1.6 63.3 D3220.1 GhomLPAT2B 27 0.9 6.6 52.230.0 7.0 0.7 1.4 61.9 D3221.39 20 0.9 6.7 53.9 28.7 6.7 0.6 1.5 63.7D3221.40 22 0.8 6.5 53.7 29.1 6.8 0.6 1.4 63.2 S6573.6 14 0.8 6.3 54.030.2 5.5 0.6 1.6 63.4 D3223.2 GhomLPAT2C 20 0.8 6.5 53.0 29.3 7.3 0.61.5 62.4 D3223.6 21 0.8 6.5 53.5 29.3 7.0 0.6 1.4 62.7 D3223.7 21 0.86.4 52.5 30.7 6.6 0.5 1.5 61.8 D3225.5 GindLPAT2A 13 0.9 6.6 53.5 30.25.6 0.6 1.6 63.2 S6573.7 12 0.9 6.5 53.5 29.9 5.7 0.6 1.8 63.3 D3227.6GindLPAT2B 23 0.8 6.4 54.1 28.8 6.8 0.6 1.6 63.5 D3227.3 21 0.8 6.5 53.929.0 6.7 0.6 1.5 63.4 D3227.17 22 0.8 6.6 53.8 28.8 7.0 0.6 1.4 63.3S6573.8 11 0.8 6.4 54.3 30.1 5.4 0.6 1.7 63.8 D3229.41 GindLPAT2C 11 0.96.6 54.2 29.7 5.6 0.6 1.7 63.9 D3229.27 13 0.8 6.4 54.1 30.0 5.6 0.6 1.763.6 D3229.33 12 0.8 6.4 54.0 30.2 5.5 0.6 1.7 63.5

Table 11 presents the TAG composition of the lipids produced by D2971,D2973, D2975, D3221, D3223, and D3227 primary transformants relative tothe S6573 parent. SOS levels in the LPAT2-expressing strains wereequivalent or slightly higher than in the S6573 controls. Trisaturatesdeclined by up to 53%, and total Sat-Unsat-Sat levels improved in all ofthe strains expressing heterologous LPAT2 genes. Among the LPAT2 genes,the strains expressing the T. cacao LPAT2 homolog showed the greatestimprovements in their TAG profiles).

TABLE 11 TAG composition of D2971, D2973, D2975, D3221, D3223, and D3227primary transformants relative to the S6573 parent. LPAAT gene BnLPAT2BnLPAT2 Ghom Ghom Gind (1.13) (1.5) TcLPAT2 LPAT2B LPAT2C LPAT2B StrainD2971.1 D2973.38 D2975.33 D2975.13 D3221.39 D3221.40 D3223.6 D3227.3D3227.6 % S6573 SOS 100 100 110 104 107 107 108 103 105 TAG Sat-Sat-Sat57 63 48 47 74 62 68 62 70 Sat-U-Sat 109 107 113 110 112 112 109 108 107Sat-O-Sat 97 100 105 102 106 105 102 104 104 Sat-L-Sat 174 147 155 155139 143 141 130 125 U-U-U/Sat 85 86 72 83 64 69 78 82 79

We analyzed the fatty acid profiles, TAG profiles and lipid titers from50 mL shake flask cultures of stable lines generated from D2975-33.C18:0 and C16:0 levels were comparable between the strains and the S6573control, and lipid titers ranged from 75-105% of the parent strain titer(Table 12). C18:2 levels increased by more than 2% in theTcLPAT2-expressing strains.

TABLE 12 Fatty acid profiles of TcLPAT2-expressing stable lines madefrom D2975-33. Primary D1940.19 D2975.33 Strain S6573 S7813 S7815 S7816S7817 S7819 Fatty Acid C12:0 0.2 0.2 0.2 0.2 0.2 0.2 Area % C14:0 0.90.7 0.8 0.8 0.7 0.7 C16:0 6.5 5.9 6.1 5.9 6.1 6.0 C16:1 0.1 0.1 0.1 0.10.1 0.1 cis-9 C17:0 0.2 0.2 0.2 0.2 0.2 0.2 C18:0 56.1 55.6 55.9 56.253.9 53.9 C18:1 28.1 26.8 26.6 26.5 28.8 28.4 C18:2 5.5 8.1 7.7 7.9 7.77.8 C18:3 α 0.6 0.5 0.6 0.5 0.6 0.7 C20:0 1.5 1.5 1.4 1.3 1.3 1.5 C22:00.2 0.2 0.1 0.1 0.1 0.2 C24:0 0.1 0.1 0.1 0.1 0.1 0.1 saturates 65.764.4 65.0 64.9 62.8 62.9

The TAG profiles of S6573 and S7815 are compared in FIG. 1. SOS levelsin the LPAT2-expressing strains were higher than in the S6573 control.Trisaturates were reduced from 10.2% in S6573 to 5.6% in S7815. Much ofthe improvement in total sat-unsat-sat levels in S7815 came from a 4%increase in stearate-linoleate-stearate (SLS) and a 1.5% increase inpalmitate-linoleate-stearate (PLS), consistent with the enhanced C18:2content of that strain. These results indicate that the T. cacoa LPAT2reduces the incorporation of saturated fatty acids at the sn-2 position.

The performance of S7815 versus the S6573 parent strain was compared inhigh-density fermentations. The fatty acid profile of each strain at thetwo time points of the fermentations are shown in Table 13. The strainshad very similar composition, with 5.5-5.7% C16:0, 56.4-56.8% C18:0, and27.2-28.6% C18:1 as the major fatty acids. As was observed in the shakeflask assays, (see Table 12), C18:2 levels increased from 5.5% in S6573to 7.7% in S7815(Table 13). Normalized lipid titers and yields werecomparable between the two strains, indicating that expression of theTcLPAT2 gene in S7815 did not have deleterious effects on growth orlipid accumulation.

TABLE 13 Fatty acid profiles of S7815 versus S6573 fermentations. StrainS6573 S7815 Fermentation 140207F25 140208F26 Fatty Acid C12:0 0.19 0.200.20 0.21 Area % C14:0 0.71 0.72 0.66 0.66 C16:0 5.69 5.73 5.57 5.54C16:1 cis-7 0.05 0.05 0.05 0.06 C16:1 cis-9 0.07 0.06 0.05 0.05 C17:00.11 0.11 0.12 0.11 C8:0 56.01 56.78 55.50 56.37 C8:1 29.31 28.58 27.9227.19 C8:2 5.56 5.51 7.75 7.70 C8:3 α 0.34 0.32 0.40 0.37 C20:0 1.511.50 1.35 1.34 C22:0 0.16 0.16 0.14 0.14 C24:0 0.10 0.09 0.09 0.08 sumC18 91.22 91.19 91.57 91.63 saturates 64.54 65.34 63.69 64.51unsaturates 35.46 34.64 36.30 35.49

Table 13 compares the TAG profiles of the lipids produced duringhigh-density fermentation of S7815 versus S6573. SOS and Sat-Oleate-Satlevels were almost identical between S7815 and the S6573 control.However, Sat-Linoleate-Sat levels increased by more than 7%, anddi-unsaturated and tri-unsaturated TAGs (U—U-U/Sat) declined by morethan 3% in S7815 compared to S6573. Trisaturates at the end points ofthe fermentations were reduced from 10.1% in S6573 to 6.1% in S7815.These results indicate that the activity of T. cacoa LPAT2 drives thetransfer of unsaturated fatty acids towards the sn-2 position anddiscriminates against the incorporation of saturated fatty acids atsn-2.

Example 6: Identification and Expression of Novel LPAAT, GPAT, DGAT,LPCAT and PLA2 with Specificity for Mid-Chain Fatty Acids

In this example, we demonstrate the effect of expression of LPAAT, GPAT,DGAT, LPCAT and PLA2 enzymes involved in triacylglycerol biosynthesis(in previously described P. moriformis (UTEX 1435) transgenic strains,S7858 and S8174. S7858 and S8174 were prepared according to co-ownedWO2015/051319, herein incorporated by reference. In addition co-ownedWO2010/063031 and WO2010/063032 teach the expression Cuphea hookerianasFATB2. Briefly, strain S7858 is a strain that express sucrose invertaseand a Cuphea. hookeriana FATB2. To make S7858, the construct pSZ4329(SEQ ID NO: 197) was engineered into S3150, a strain classicallymutagenized to increase lipid yield. The plasmid, pSZ4329 is written asTHI4α::CrTUB2-ScSUC2-PmPGH:PmAcp-P1p-CpSAD1tp_trimmed_ChFATB2_FLAG-CvNR::THI4aThe annotation of the coding portions of pSZ4329 is shown in the Table Abelow.

TABLE A Nucleotide Nucleotide Nucleotide pSZ4329 Identity Number NumberLength THI4a 3′ flank 3′ flanking sequences 5,692 6,394 703 ofendogenous THI4 CvNR 3′UTR 5,278 5,679 402 ChFATB2 CDS 4,105 5,271 1,167CpSAD1tp-trimmed CDS 3,991 4,104 114 PmACP-P1 promoter promoter 3,4113,981 571 Buffer DNA 3,199 3,404 206 UTR04424=PmPGH 3′UTR 2,749 3,192444 UTR ScSUC2(o) CDS 1,144 2,742 1,599 CrTUB2 promoter promoter THI4a5′ flank 5′ flanking sequences 820 1,131 312 of endogenous THI4 27 813787

Strain S7858, accumulates C8:0 fatty acids to about 12% and C10:0 fattyacids to about 22-24%. Briefly, strain S8174 is a strain that expresssucrose invertase and a Cuphea. Avigera var. pulcherrima FATB2. To makeS8174, the construct pSZ5078 (SEQ ID NO: 198) was engineered into S3150,a strain classically mutagenized to increase lipid yield. pSZ5078 iswritten asTHI4a5′::CrTUB2_ScSUC2_PmPGH:PmAMT3_CpSAD1tp_trimmed-CaFATB1_Flag_CvNR::THI4a3′.Strain S8174 accumulates C8:0 fatty acids to about 24% and C10:0 fattyacids to about 10%. The annotation of the coding portions of pSZ5078 isshown in the Table B below.

TABLE B Nucleotide Nucleotide Nucleotide pSZ5078 Identity Number NumberLength THI4a 3′ 3′ flanking sequences 6,200 6,902 703 flank ofendogenous THI4 CvNR 3′UTR 5,786 6,187 402 CaFATB1 CDS 4,602 5,771 1,170wild-type CpSAD1tp CDS 4,488 4,601 114 AMT3 promoter eukaryotic 3,4114,481 1,071 Buffer DNA misc_feature 3,199 3,404 206 PmPGH 3′UTR 2,7493,192 444 ScSUC2(o) CDS 1,144 2,742 1,599 CrTUB2 promoter 820 1,131 312promoter THI4a 5′ 5′ flanking sequences 27 813 787 flank of endogenousTHI4

The pool of acyl-CoAs in the ER can be utilized for the synthesis ofTAGs as well as phospholipids and long chain fatty acids. The enzymesinvolved in the synthesis of TAGS and phospholids actively competeagainst each other for the same substrates. Acyl-CoAs can associate withlysophosphatidate to form phosphatidate which is converted tophosphatidylcholine (PC) and other phospholipid species. PC can bedesaturated by FAD2 and FAD3 enzymes to generate polyunsaturated fattyacids, which can be cleaved by phosphotransferases and reenter theacyl-CoA pool. Acyl-CoAs can also be generated from PC directly byacyl-CoA:lysophosphatidylcholine acyltransferase (LPCAT). LPCAT can alsocatalyze the reverse reaction to consume acyl-CoA. Removal of fattyacids from PC to form acyl-CoAs can also be catalyzed by phospholipaseA2 (PLA2). TAG formation in the ER from acyl-CoAs requires action ofglycerol phosphate acyltransferase (GPAT), lysophosphatidic acidacyltransferase (LPAAT) and diacyl glycerol acyltransferase (DGAT).

The endogenous P. moriformis TAG biosynthesis machinery has evolved tofunction with the longer chain fatty acids that the strain normallymakes. We introduced heterologous acyltransferases and phospholipasesfrom species that naturally accumulate high levels of short chain fattyacids into Prototheca to increase accumulation of C8:0 fatty acids. Weidentified the following plant enzymes in NCBI as shown in Table 14below.

TABLE 14 Genes representing target enzymes identified from higher plantsthat produce high amounts of C8:0 and C10:0. All these genes weresynthesized with codon usage optimized for expression in Prototheca.Species Gene Enzyme

cnLPAAT1 LPAAT Cuphea 

 LPAAT1 Cuphea 

 LPAAT1 Cuphea 

 LPAAT1 Cuphea 

 LPAAT1 Cuphea 

 LPAAT1 Cuphea avigera var. 

 LPAAT1 Cuphea avigera var. 

 LPAAT2 Cuphea 

 LPAAT1 Cuphea 

 LPAAT1 Cuphea 

 LPAAT2 Cuphea 

 LPAAT2 Cuphea 

 LPAAT2 Cuphea avigera var. 

 GPAT9 

GPAT Cuphea 

 GPAT9 

 1 Cuphea 

 GPAT9 

 2 Cuphea 

 GPAT9 

 2 Cuphea 

 GPAT9 

 2 Cuphea 

 GPAT9 

 2 Cuphea avigera var. 

 DGAT1 DGAT Cuphea 

 DGAT1 

 1 Cuphea avigera var. 

 LPCAT LPCAT Cuphea 

 LPCAT Cuphea 

 LPCAT Cuphea 

 LPCAT1 Cuphea avigera var. 

 PLA2 

 1 PLA2 Cuphea 

 PLA2 

 1 Cuphea 

 PLA2 

 2 Cuphea 

 PLA2 

 2

indicates data missing or illegible when filed

We made a set of constructs expressing heterologous short chain specificacyltransferases and PLA2s as shown in Table 15. The genes were codonoptimized to reflect UTEX 1435 codon usage.

TABLE 15 List of constructs transformed into S7858 or S8174 D# StrainConstruct D4289 S7858 SAD2-1vD::CpauLPAAT1 

 PmATP:PmHXT1 

 ScarMEL-PmPGK::SAD2Bex D4290 S7858 SAD2-1vD:: 

 LPAAT1 

 PmATP:PmHXT1 

 ScarMEL-PmPGK::SAD2Bex D4291 S7858 SAD2-1vD::CigneaLPAAT1 

 PmATP:PmHXT1 

 ScarMEL-PmPGK::SAD2Bex D4292 S7858 SAD2-1vD:: 

 LPAAT1 

 PmATP:PmHXT1 

 ScarMEL-PmPGK::SAD2Bex D4293 S7858 SAD2-1vD::ChookLPAAT1 

 PmATP:PmHXT1 

 ScarMEL-PmPGK::SAD2Bex D4404 S7858 SAD2-1vD::CnLPAAT1 

 PmATP:PmHXT1 

 ScarMEL-PmPGK::SAD2Bex D4517 S8174 SAD2-1vD::CavigLPAAT1 

 PmATP:PmHXT1 

 ScarMEL-PmPGK::SAD2Bex D4518 S8174 SAD2-1vD::CavigLPAAT2 

 PmATP:PmHXT1 

 ScarMEL-PmPGK::SAD2Bex D4519 S8174 SAD2-1vD::CavigLPAAT1 

 PmATP:PmHXT1 

 ScarMEL-PmPGK::SAD2Bex D4690 S8174 SAD2-1vD::CuPSR23 LPAAT2 

 1 

 PmATP:PmHXT1 

 ScarMEL-PmPGK::SAD2Bex D4728 S8174 SAD2-1vD::CkoeLPAAT 

 PmATP:PmHXT1 

 ScarMEL-PmPGK::SAD2Bex D4729 S8174 SAD2-1vD::CkoeLPAAT2 

 PmATP:PmHXT1 

 ScarMEL-PmPGK::SAD2Bex D4730 S8174 SAD2-1vD::CprocLPAAT2 

 PmATP:PmHXT1 

 ScarMEL-PmPGK::SAD2Bex D4551/D5683 S8174 SAD2-1vD::CavigGPAT9 

 PmATP:PmHXT1 

 ScarMEL-PmPGK::SAD2Bex D4552/D4684 S8174SAD2-1vD::ChookGPAT9-1-PmATP:PmHXT1 

 ScarMEL-PmPGK::SAD2Bex D4553/D4685 S8174SAD2-1vD::CignGPAT9-1-PmATP:PmHXT1 

 ScarMEL-PmPGK::SAD2Bex D4554/D4686 S8174SAD2-1vD::CignGPAT9-2-PmATP:PmHXT1 

 ScarMEL-PmPGK::SAD2Bex D4724 S8174 SAD2-1vD:: 

 GPAT9-1-PmATP:PmHXT1 

 ScarMEL-PmPGK::SAD2Bex D4725 S8174 SAD2-1vD:: 

 GPAT9-2-PmATP:PmHXT1 

 ScarMEL-PmPGK::SAD2Bex D4549 S8174 SAD2-1vD::CavigDGAT1 

 PmATP:PmHXT1 

 ScarMEL-PmPGK::SAD2Bex D4681 S8174 SAD2-1vD::CavigDGAT1 

 PmATP:PmHXT1 

 ScarMEL-PmPGK::SAD2Bex D4556/D4688 S8174 SAD2-1vD::CavigLPCAT 

 PmATP:PmHXT1 

 ScarMEL-PmPGK::SAD2Bex D4726 S8174 SAD2-1vD:: 

 LPCAT-PmATP:PmHXT1 

 ScarMEL-PmPGK::SAD2Bex D4556/D4689 S8174SAD2-1vD::CpauLPCAT-PmATP:PmHXT1 

 ScarMEL-PmPGK::SAD2Bex D4727 S8174 SAD2-1vD::CschuLPCAT-PmATP:PmHXT1 

 ScarMEL-PmPGK::SAD2Bex D4732 S8174 SAD2-1vD::CavigPLA2-1-PmATP:PmHXT1 

 ScarMEL-PmPGK::SAD2Bex D4734 S8174 SAD2-1vD::CignPLA2-1-PmATP:PmHXT1 

 ScarMEL-PmPGK::SAD2Bex D4735 S8174SAD2-1vD::CuPSR23PLA2-2-PmATP:PmHXT1 

 ScarMEL-PmPGK::SAD2Bex D4736 S8174 SAD2-1vD::CprocPLA2-2-PmATP:PmHXT1 

 ScarMEL-PmPGK::SAD2Bex

indicates data missing or illegible when filed

All the constructs shown in Table 15 can be written as SAD2-1vD::gene ofinterest-PmATP-PmHXT1-ScarMEL-PmPGK::SAD2B, and were made to target thetransforming DNA to the SAD2 locus on the genome, thereby disrupting theexpression of at least one allele of the endogenous stearoyl ACPdesaturase. Sequences of all the transforming DNAs are provided below.The relevant restriction sites in the construct from 5′-3′ are- Pme I,BspQ I, Kpn I, Xho I, Avr II, Spe I, SnaB I, EcoR V, Sac I, BspQ I, PmeI respectively are indicated in lowercase, bold, and underlined. Pme Isites delimit the 5′ and 3′ ends of the transforming DNA. Bold,lowercase sequences at the 5′ and 3′ end of the construct representgenomic DNA from UTEX 1435 that target integration to the SAD2 locus viahomologous recombination, wherein the SAD2 5′ flank provides thepromoter for the gene of interest downstream. The primary construct wasmade with the previously characterized CnLPAAT gene as shown below andall other constructs were made by replacing the CnLPAAT gene with othergenes of interest using the restriction sites, Kpn I and Xho I that spanthe gene on either side. Proceeding in the 5′ to 3′ direction, the firstcassette has the codon optimized Cocos nucifera LPAAT and the Protothecamoriformis ATP synthase (PmATP) gene 3′ UTR. The initiator ATG andterminator TGA for cDNAs are indicated by uppercase italics, while thecoding region is indicated with lowercase italics. The 3′ UTR isindicated by lowercase underlined text. The second cassette containingthe selection gene melibiose from Saccharomyces carlsbergensis(ScarMEL1) is driven by the endogenous HXT1 promoter, and has theendogenous phosphoglycerate kinase (PmPGK) gene 3′ UTR. In thiscassette, the PmHXT1 promoter is indicated by lowercase, boxed text. Theinitiator ATG and terminator TGA for the ScarMEL1 gene are indicated inuppercase italics, while the coding region is indicated by lowercaseitalics. The 3′ UTR is indicated by lowercase underlined text. All thefinal constructs were sequenced to ensure correct reading frames andtargeting sequences.

pSZX61 Sequence of the transforming DNA expressingCnLPAAT downstream of the SAD2 promoter in the cassette followed by the ScarMEL1gene for selection downstream of the PmHXT1 promoter in the second cassette.SEQ ID NO: 97 gtttaaacgccggtcaccacccgcatgctcgtactacagcgcacgcaccgcttcgtgatccaccgggtgaacgtagtcctcgacggaaacatctggttcgggcctcctgcttgcactcccgcccatgccgacaacctttctgctgttaccacgacccacaatgcaacgcgacacgaccgtgtgggactgatcggttcactgcacctgcatgcaattgtcacaagcgcttactccaattgtattcgtttgttttctgggagcagttgctcgaccgcccgcgtcccgcaggcagcgatgacgtgtgcgtggcctgggtgtttcgtcgaaaggccagcaaccctaaatcgcaggcgatccggagattgggatctgatccgagtttggaccagatccgccccgatgcggcacgggaactgcatcgactcggcgcggaacccagctttcgtaaatgccagattggtgtccgatacctggatttgccatcagcgaaacaagacttcagcagcgagcgtatttggcgggcgtgctaccagggttgcatacattgcccatttctgtctggaccgctttactggcgcagagggtgagttgatggggttggcaggcatcgaaacgcgcgtgcatggtgtgcgtgtctgttttcggctgcacgaattcaatagtcggatgggcgacggtagaattgggtgtggcgctcgcgtgcatgcctcgccccgtcgggtgtcatgaccgggactggaatcccccctcgcgaccatcttgctaacgctcccgactctcccgaccgcgcgcaggatagactcttgttcaaccaatcgaca ggtaccATGgacgcctccggcgcctcctccttcctgcgcggccgctgcctggagtcctgcttcaaggcctccttcggctacgtaatgtcccagcccaaggacgccgccggccagccctcccgccgccccgccgacgccgacgacttcgtggacgacgaccgctggatcaccgtgatcctgtccgtggtgcgcatcgccgcctgcttcctgtccatgatggtgaccaccatcgtgtggaacatgatcatgctgatcctgctgccctggccctacgcccgcatccgccagggcaacctgtacggccacgtgaccggccgcatgctgatgtggattctgggcaaccccatcaccatcgagggctccgagttctccaacacccgcgccatctacatctgcaaccacgcctccctggtggacatcttcctgatcatgtggctgatccccaagggcaccgtgaccatcgccaagaaggagatcatctggtatcccctgttcggccagctgtacgtgctggccaaccaccagcgcatcgaccgctccaacccctccgccgccatcgagtccatcaaggaggtggcccgcgccgtggtgaagaagaacctgtccctgatcatcttccccgagggcacccgctccaagaccggccgcctgctgcccttcaagaagggcttcatccacatcgccctccagacccgcctgcccatcgtgccgatggtgctgaccggcacccacctggcctggcgcaagaactccctgcgcgtgcgccccgcccccatcaccgtgaagtacttctcccccatcaagaccgacgactgggaggaggagaagatcaaccactacgtggagatgatccacgccctgtacgtggaccacctgcccgagtcccagaagcccctggtgtccaagggccgcgacgcctccggccgctccaactccTGAttaattaactcga g atgtggagatgtagggtggtcgactcgttggaggtgggtgtttttttttatcgagtgcgcggcgcggcaaacgggtccctttttatcgaggtgttcccaacgccgcaccgccctcttaaaacaacccccaccaccacttgtcgaccttctcgtttgttatccgccacggcgccccggaggggcgtcgtctggccgcgcgggcagctgtatcgccgcgctcgctccaatggtgtgtaatcttggaaagataataatcgatggatgaggaggagagcgtgggagatcagagcaaggaatatacagttggcacgaagcagcagcgtactaagctgtagcgtgttaagaaagaaaaactcg

cgtctccccctcctacaacggcctgggcctgacgccccagatgggctgggacaactggaacacgttcgcctgcgacgtctccgagcagctgctgctggacacggccgaccgcatctccgacctgggcctgaaggacatgggctacaagtacatcatcctggacgactgctggtcctccggccgcgactccgacggcttcctggtcgccgacgagcagaagaccccaacggcatgggccacgtcgccgaccacctgcacaacaactccttcctgacggcatgtactcctccgcgggcgagtacacgtgcgccggctaccccggctccctgggccgcgaggaggaggacgcccagttcttcgcgaacaaccgcgtggactacctgaagtacgacaactgctacaacaagggccagttcggcacgcccgagatctcctaccaccgctacaaggccatgtccgacgccctgaacaagacgggccgccccatcttctactccctgtgcaactggggccaggacctgaccttctactggggctccggcatcgcgaactcctggcgcatgtccggcgacgtcacggcggagttcacgcgccccgactcccgctgcccctgcgacggcgacgagtacgactgcaagtacgccggcttccactgctccatcatgaacatcctgaacaaggccgcccccatgggccagaacgcgggcgtcggcggctggaacgacctggacaacctggaggtcggcgtcggcaacctgacggacgacgaggagaaggcgcacttctccatgtgggccatggtgaagtcccccctgatcatcggcgcgaacgtgaacaacctgaaggcctcctcctactccatctactcccaggcgtccgtcatcgccatcaaccaggactccaacggcatccccgccacgcgcgtctggcgctactacgtgtccgacacggacgagtacggccagggcgagatccagatgtggtccggccccctggacaacggcgaccaggtcgtggcgctgctgaacggcggctccgtgtcccgccccatgaacacgaccctggaggagatcttcttcgactccaacctgggctccaagaagctgacctccacctgggacatctacgacctgtgggcgaaccgcgtcgacaactccacggcgtccgccatcctgggccgcaacaagaccgccaccggcatcctgtacaacgccaccgagcagtcctacaaggacggcctgtccaagaacgacacccgcctgttcggccagaagatcggctccctgtcccccaacgcgatcctgaacacgaccgtccccgcccacggcatcgcgttctaccgcctgcgcccctcctccTGA tacaacttattacgtattctgaccggcgctgatgtggcgcggacgccgtcgtactctttcagactttactcttgaggaattgaacctttctcgcttgctggcatgtaaacattggcgcaattaattgtgtgatgaagaaagggtggcacaagatggatcgcgaatgtacgagatcgacaacgatggtgattgttatgaggggccaaacctggctcaatcttgtcgcatgtccggcgcaatgtgatccagcggcgtgactctcgcaacctggtagtgtgtgcgcaccgggtcgctttgattaaaactgatcgcattgccatcccgtcaactcacaagcctactctagctcccattgcgcactcgggcgcccggctcgatcaatgttctgagcggagggcgaagcgtcaggaaatcgtctcggcagctggaagcgcatggaatgcggagcggagatcgaatcagatatcAAGCTCCATC gagctc cagccacggcaacaccgcgcgccttgcggccgagcacggcgacaagaacctgagcaagatctgcgggctgatcgccagcgacgagggccggcacgagatcgcctacacgcgcatcgtggacgagttcttccgcctcgaccccgagggcgccgtcgccgcctacgccaacatgatgcgcaagcagatcaccatgcccgcgcacctcatggacgacatgggccacggcgaggccaacccgggccgcaacctcttcgccgacttctccgcggtcgccgagaagatcgacgtctacgacgccgaggactactgccgcatcctggagcacctcaacgcgcgctggaaggtggacgagcgccaggtcagcggccaggccgccgcggaccaggagtacgtcctgggcctgccccagcgcttccggaaactcgccgagaagaccgccgccaagcgcaagcgcgtcgcgcgcaggcccgtcgccttctcctggatctccgggcgcgagatcatggtctagggagcgacgagtgtgcgtgcggggctggcgggagtgggacgccctcctcgctcctctctgttctgaacggaacaatcggccaccccgcgctacgcgccacgcatcgagcaacgaagaaaaccccccgatgataggttgcggtggctgccgggatatagatccggccgcacatcaaagggcccctccgccagagaagaagctcctttcccagcagactcct gaagagcgtttaaac .

The sequence for all of the other acyltransferase constructs areidentical to that of pSZEX61 with the exception of the encodedacyltransferase. The acyltransferase sequence alone is provided belowfor the remaining acyltransferase constructs.

SEQ ID NO: 98 CpauLPAAT1 ggtaccATGgccatccccgccgccgccgtgatcttcctgttcggcctgctgttcttcacctccggcctgatcatcaacctgttccaggccctgtgcttcgtgctggtgtggcccctgtccaagaacgcctaccgccgcatcaaccgcgtgttcgccgagctgctgctgtccgagctgctgtgcctgttcgactggtgggccggcgccaagctgaagctgttcaccgaccccgagaccttccgcctgatgggcaaggagcacgccctggtgatcatcaaccacatgaccgagctggactggatgctgggctgggtgatgggccagcacctgggctgcctgggctccatcctgtccgtggccaagaagtccaccaagttcctgcccgtgctgggctggtccatgtggttctccgagtacctgtacatcgagcgctcctgggccaaggaccgcaccaccctgaagtcccacatcgagcgcctgaccgactaccccctgcccttctggatggtgatcttcgtggagggcacccgcttcacccgcaccaagctgctggccgcccagcagtacgccgcctcctccggcctgcccgtgccccgcaacgtgctgatcccccgcaccaagggcttcgtgtcctgcgtgtcccacatgcgctccttcgtgcccgccgtgtacgacgtgaccgtggccttccccaagacctcccccccccccaccctgctgaacctgttcgagggccagtccatcgtgctgcacgtgcacatcaagcgccacgccatgaaggacctgcccgagtccgacgacgccgtggcccagtggtgccgcgacaagttcgtggagaaggacgccctgctggacaagcacaacgccgaggacaccttctccggccaggaggtgcaccgcaccggctcccgccccatcaagtccctgctggtggtgatctcctgggtggtggtgatcaccttcggcgccctgaagttcctgcagtggtcctcctggaagggcaaggccttctccgtgatcggcctgggcatcgtgaccctgctgatgcacatgctgatcctgtcctcccaggccgagcgctcctccaaccccgccaaggtggcccaggccaagctgaagaccgagctgtccatctccaagaaggccaccgacaaggagaacTGA ctcgagSEQ ID NO: 99 CprocLPAAT1 ggtacc

ctcgag SEQ ID NO: 100 CpaiLPAAT1 ggtaccATGgccatcccctccgccgccgtggtgacctgacggcctgctgacttcacctccggcctgatcatcaacctgaccaggccactgcacgtgctgatctcccccctgtccaagaacgcctaccgccgcatcaaccgcgtgacgccgagctgctgcccctggagttcctgtggctgaccactggtgcgccggcgccaagctgaagctgacaccgaccccgagaccaccgcctgatgggcaaggagcacgccctggtgatcatcaaccacaagatcgagctggactggatggtgggctgggtgctgggccagcacctgggctgcctgggctccatcctgtccgtggccaagaagtccaccaagacctgcccgtgacggctggtccctgtggactccggctacctgacctggagcgctcctgggccaaggacaagatcaccctgaagtcccacatcgagtccctgaaggactaccccctgccataggctgatcatcacgtggagggcacccgatcacccgcaccaagctgctggccgcccagcagtacgccgcctcctccggcctgcccgtgccccgcaacgtgctgatcccccacaccaagggcttcgtgtcctccgtgtcccacatgcgctccttcgtgcccgccatctacgacgtgaccgtggccttccccaagacctcccccccccccaccatgctgaagctgacgagggccagtccgtggagctgcacgtgcacatcaagcgccacgccatgaaggacctgcccgagtccgacgacgccgtggcccagtggtgccgcgacaagacgtggagaaggacgccctgctggacaagcacaactccgaggacaccactccggccaggaggtgcaccacgtgggccgccccatcaaggccctgctggtggtgatctcctgggtggtggtgatcatcacggcgccctgaagacctgctgtggtcctccctgctgtcctcctggaagggcaaggccactccgtgatcggcctgggcatcgtggccggcatcgtgaccctgctgatgcacatcctgatcctgtcctcccaggccgagggctccaaccccgtgaaggccgcccccgccaagctgaagaccgagctgtcctcctccaagaaggtgaccaacaaggagaacTGA ctcgagSEQ ID NO: 101 ChookLPAAT1 ggtaccATGgccatcccctccgccgccgtggtgacctgacggcctgctgacttcacctccggcctgatcatcaacctgaccaggccactgatcgtgctgatctcccccctgtccaagaacgcctaccgccgcatcaaccgcgtgacgccgagctgctgcccctggagttcctgtggctgaccactggtgcgccggcgccaagctgaagctgacaccgaccccgagaccaccgcctgatgggcaaggagcacgccctggtgatcatcaaccacaagatcgagctggactggatggtgggctgggtgctgggccagcacctgggctgcctgggctccatcctgtccgtggccaagaagtccaccaagacctgcccgtgacggctggtccctgtggactccgagtacctgacctggagcgctcctgggccaaggacaagatcaccctgaagtcccacatcgagtccctgaaggactaccccctgccataggctgatcatcacgtggagggcacccgatcacccgcaccaagctgctggccgcccagcagtacgccgcctcctccggcctgcccgtgccccgcaacgtgctgatcccccacaccaagggcttcgtgtcctccgtgtcccacatgcgctccttcgtgcccgccatctacgacgtgaccgtggccttccccaagacctcccccccccccaccatgctgaagctgacgagggccagtccgtggagctgcacgtgcacatcaagcgccacgccatgaaggacctgcccgagtccgacgacgccgtggcccagtggtgccgcgacaagacgtggagaaggacgccctgctggacaagcacaactccgaggacaccactccggccaggaggtgcaccacgtgggccgccccatcaaggccctgctggtggtgatctcctgggtggtggtgatcatcacggcgccctgaagacctgctgtggtcctccctgctgtcctcctggaagggcaaggccactccgtgatcggcctgggcatcgtggccggcatcgtgaccctgctgatgcacatcctgatcctgtcctcccaggccgagggctccaaccccgtgaaggccgcccccgccaagctgaagaccgagctgtcctcctccaagaaggtgaccaacaaggagaacTGA ctcgagSEQ ID NO: 102 CignLPAAT1 ggtaccATGgccatcgccgccgccgccgtgatcacctgacggcctgctgacttcgcctccggcatcatcatcaacctgaccaggccctgtgcttcgtgctgatctggcccctgtccaagaacgtgtaccgccgcatcaaccgcgtgttcgccgagctgctgctgatggacctgctgtgcctgaccactggtgggccggcgccaagatcaagctgacaccgaccccgagaccaccgcctgatgggcatggagcacgccctggtgatcatgaaccacaagaccgacctggactggatggtgggctggatcctgggccagcacctgggctgcctgggctccatcctgtccatcgccaagaagtccaccaagacatccccgtgctgggctggtccgtgtggactccgagtacctgacctggagcgctcctgggccaaggacaagtccaccctgaagtcccacatggagaagctgaaggactaccccctgccataggctggtgatcacgtggagggcacccgatcacccgcaccaagctgctggccgcccagcagtacgccgcctcctccggcctgcccgtgccccgcaacgtgctgatcccccacaccaagggcttcgtgtcctgcgtgtccaacatgcgctccttcgtgcccgccgtgtacgacgtgaccgtggccttccccaagtcctcccccccccccaccatgctgaagctgacgagggccagtccatcgtgctgcacgtgcacatcaagcgccacgccctgaaggacctgcccgagtccgacgacgccgtggcccagtggtgccgcgacaagacgtggagaaggacgccctgctggacaagcacaacgccgaggacaccactccggccaggaggtgcaccacatcggccgccccatcaagtccctgctggtggtgatcgcctgggtggtggtgatcatcacggcgccctgaagacctgcagtggtcctccctgctgtccacctggaagggcaaggccactccgtgatcggcctgggcatcgccaccctgctgatgcacatgctgatcctgtcctcccaggccgagcgctccaaccccgccaaggtggccaagTGA ctcgag SEQ ID NO: 103 CavigLPAAT1 ggtaccATGaccatcgcctccgccgccgtggtgttcctgttcggcatcctgctgttcacctccggcctgatcatcaacctgttccaggccttctgctccgtgctggtgtggcccctgtccaagaacgcctaccgccgcatcaaccgcgtgttcgccgagttcctgcccctggagttcctgtggctgttccactggtgggccggcgccaagctgaagctgttcaccgaccccgagaccttccgcctgatgggcaaggagcacgccctggtgatcatcaaccacaagatcgagctggactggatggtgggctgggtgctgggccagcacctgggctgcctgggctccatcctgtccgtggccaagaagtccaccaagacctgcccgtgttcggctggtccctgtggttctccgagtacctgttcctggagcgcaactgggccaaggacaagaagaccctgaagtcccacatcgagcgcctgaaggactaccccctgcccttctggctgatcatcttcgtggagggcacccgcttcacccgcaccaagctgctggccgcccagcagtacgccgcctccgccggcctgcccgtgccccgcaacgtgctgatcccccacaccaagggatcgtgtcctccgtgtcccacatgcgctccacgtgcccgccatctacgacgtgaccgtggccttccccaagacctcccccccccccaccatgctgaagctgttcgagggccacttcgtggagctgcacgtgcacatcaagcgccacgccatgaaggacctgcccgagtccgaggacgccgtggcccagtggtgccgcgacaagttcgtggagaaggacgccctgctggacaagcacaacgccgaggacaccttctccggccaggaggtgcaccacgtgggccgccccatcaagtccctgctggtggtgatctcctgggtggtggtgatcatcttcggcgccctgaagttcctgcagtggtcctccctgctgtcctcctggaagggcatcgccttctccgtgatcggcctgggcaccgtggccctgctgatgcagatcctgatcctgtcctcccaggccgagcgctccatccccgccaaggagacccccgccaacctgaagaccgagctgtcctcctccaagaaggtgaccaacaaggagaacTGA ctcgagSEQ ID NO: 104 CavigLPAAT2 ggtaccATGgccatcgccgccgccgccgtgatcgtgcccgtgtccctgctgttcttcgtgtccggcctgatcgtgaacctggtgcaggccgtgtgcttcgtgctgatccgccccctgttcaagaacacctaccgccgcatcaaccgcgtggtggccgagctgctgtggctggagctggtgtggctgatcgactggtgggccggcgtgaagatcaaggtgttcaccgaccacgagaccttccacctgatgggcaaggagcacgccctggtgatctgcaaccacaagtccgacatcgactggctggtgggctgggtgctggcccagcgctccggctgcctgggctccaccctggccgtgatgaagaagtcctccaagttcctgcccgtgatcggctggtccatgtggactccgagtacctgttcctggagcgcaactgggccaaggacgagtccaccctgaagtccggcctgaaccgcctgaaggactaccccctgcccttctggctggccctgttcgtggagggcacccgcttcacccgcgccaagctgctggccgcccagcagtacgccgcctcctccggcctgcccgtgccccgcaacgtgctgatcccccgcaccaagggatcgtgtcctccgtgtcccacatgcgctcatcgtgcccgccatctacgacgtgaccgtggccatccccaagacctcccccccccccaccctgctgcgcatgttcaagggccagtcctccgtgctgcacgtgcacctgaagcgccaccagatgaacgacctgcccgagtccgacgacgccgtggcccagtggtgccgcgacatcttcgtggagaaggacgccctgctggacaagcacaacgccgaggacaccttctccggccaggagctgcaggacaccggccgccccatcaagtccctgctgatcgtgatctcctgggccgtgctggtggtgttcggcgccgtgaagttcctgcagtggtcctccctgctgtcctcctggaagggcctggccttctccggcatcggcctgggcgtgatcaccctgctgatgcacatcctgatcctgttctcccagtccgagcgctccacccccgccaaggtggcccccgccaagcccaagatcgagggcgagtcctccaagaccgagatggagaaggagcacTGA ctcgagSEQ ID NO: 105 CpalLPAAT1 ggtaccATGgccatcgccgccgccgccgtgatcgtgcccctgggcctgctgttcttcgtgtccggcctgatcgtgaacctggtgcaggccgtgtgcttcgtgctgatccgccccctgtccaagaacacctaccgccgcatcaaccgcgtggtggccgagctgctgtggctggagctggtgtggctgatcgactggtgggccggcgtgaagatcaaggtgttcaccgaccacgagaccctgtccctgatgggcaaggagcacgccctggtgatctgcaaccacaagtccgacatcgactggctggtgggctgggtgctggcccagcgctccggctgcctgggctccaccctggccgtgatgaagaagtcctccaagttcctgcccgtgatcggctggtccatgtggttctccgagtacctgcccgagtccgacgacgccgtggcccagtggtgccgcgacatcttcgtggagaaggacgccctgctggacaagcacaacgccgaggacaccttctccggccaggagctgcaggacaccggccgccccatcaagtccctgctggtggtgatctcctgggccgtgctggtgatcttcggcgccgtgaagttcctgcagtggtcctccctgctgtcctcctggaagggcctggccttctccggcgtgggcctgggcatcatcaccctgctgatgcacatcctgatcctgttctcccagtccgagcgctccacccccgccaaggtggcccccgccaagcccaagaaggacggcgagtcctccaagaccgagatcgagaaggagaacgttcctggagcgctcctgggccaaggacgagaacaccctgaagtccggcctgaaccgcctgaaggactaccccctgcccttctggctggccctgttcgtggagggcacccgcttcacccgcgccaagctgctggccgcccagcagtacgccacctcctccggcctgcccgtgccccgcaacgtgctgatcccccgcaccaagggcttcgtgtcctccgtgtcccacatgcgctcatcgtgcccgccatctacgacgtgaccgtggccatccccaagacctcccccccccccaccatgctgcgcatgttcaagggccagtcctccgtgctgcacgtgcacctgaagcgccacctgatgaaggacctTGA ctcgagSEQ ID NO: 106 CuPSR23 LPAAT2 ggtaccATGgccatcgccgccgccgccgtgatcacctgttcggcctgatatatcgcctccggcctgatcatcaacctgttccaggccctgtgcttcgtgctgatccgccccctgtccaagaacgcctaccgccgcatcaaccgcgtgttcgccgagctgctgctgtccgagctgctgtgcctgacgactggtgggccggcgccaagctgaagctgttcaccgaccccgagaccaccgcctgatgggcaaggagcacgccctggtgatcatcaaccacatgaccgagctggactggatggtgggctgggtgatgggccagcacttcggctgcctgggctccatcatctccgtggccaagaagtccaccaagacctgcccgtgctgggctggtccatgtggactccgagtacctgtacctggagcgctcctgggccaaggacaagtccaccctgaagtcccacatcgagcgcctgatcgactaccccctgcccactggctggtgatcacgtggagggcacccgcttcacccgcaccaagctgctggccgcccagcagtacgccgtgtcctccggcctgcccgtgccccgcaacgtgctgatcccccgcaccaagggcacgtgtcctgcgtgtcccacatgcgctccacgtgcccgccgtgtacgacgtgaccgtggccaccccaagacctcccccccccccaccctgctgaacctgacgagggccagtccatcatgctgcacgtgcacatcaagcgccacgccatgaaggacctgcccgagtccgacgacgccgtggccgagtggtgccgcgacaagacgtggagaaggacgccctgctggacaagcacaacgccgaggacaccactccggccaggaggtgtgccactccggctcccgccagctgaagtccctgctggtggtgatctcctgggtggtggtgaccaccttcggcgccctgaagacctgcagtggtcctcctggaagggcaaggccactccgccatcggcctgggcatcgtgaccctgctgatgcacgtgctgatcctgtcctcccaggccgagcgctccaaccccgccgaggtggcccaggccaagctgaagaccggcctgtccatctccaagaaggtgaccgacaaggagaacTGA ctcgagSEQ ID NO: 107 CkoeLPAAT1 ggtaccATGgccatccccgccgccgtggccgtgatccccatcggcctgctgacatcatctccggcctgatcgtgaacctgatccaggccgtggtgtacgtgctgatccgccccctgtccaagaacctgcaccgcaagatcaacaagcccatcgccgagctgctgtggctggagctgatctggctggtggactggtgggccggcatcaaggtggaggtgtacgccgactcccagaccctggagctgatgggcaaggagcacgccctgctgatctgcaaccaccgctccgacatcgactggctggtgggctgggtgctggcccagcgcgcccgctgcctgggctccgccctggccatcatgaagaagtccgccaagttcctgcccgtgatcggctggtccatgtggactccgactacatcacctggaccgcacctgggccaaggacgagaagaccctgaagtccggatcgagcgcctggccgacttccccatgccatctggctggccctgacgtggagggcacccgatcaccaaggccaagctgctggccgcccaggagtacgccgcctcccgcggcctgcccgtgccccagaacgtgctgatcccccgcaccaagggatcgtgaccgccgtgacccacatgcgctcctacgtgcccgccatctacgactgcaccgtggacatctccaaggcccaccccgccccctccatcctgcgcctgatccgcggccagtcctccgtggtgaaggtgcagatcacccgccactccatgcaggagctgcccgagaccgccgacggcatctcccagtggtgcatggacctgttcgtgaccaaggacggcacctggagaagtaccactccaaggacatcacggctccctgcccgtgcagaacatcggccgccccgtgaagtccctgatcgtggtgctgtgctggtactgcctgatggccacggcctgacaagacttcatgtggtcctccctgctgtcctcctgggagggcatcctgtccctgggcctgatcctgctggccgtggccatcgtgatgcagatcctgatccagtccaccgagtccgagcgctccacccccgtgaagtccatccagaaggacccctccaaggagaccctgctgcagaacTGA ctcgag SEQ ID NO: 108 CkoeLPAAT2ggtaccATGcacgtgctgctggagatggtgaccaccgcactcctccacttcgtgacgacaacgtgcaggccctgtgatcgtgctgatctggcccctgtccaagtccgcctaccgcaagatcaaccgcgtgacgccgagctgctgctgtccgagctgctgtgcctgacgactggtgggccggcgccaagctgaagctgacaccgaccccgagaccaccgcctgatgggcaaggagcacgccctggtgatcaccaaccacaagatcgacctggactggatgatcggctggatcctgggccagcacttcggctgcctgggctccgtgatctccatcgccaagaagtccaccaagacctgcccatcacggctggtccctgtggactccgagtacctgacctggagcgcaactgggccaaggacaagcgcaccctgaagtcccacatcgagcgcatgaaggactaccccctgcccctgtggctgatcctgacgtggagggcacccgatcacccgcaccaagctgctggccgcccagcagtacgccgcctcctccggcctgcccgtgccccgcaacgtgctgatcccccacaccaagggcttcgtgtcctccgtgtcccacatgcgctccttcgtgcccgccgtgtacgacgtgaccgtggccttccccaagacctcccccccccccaccatgctgtccctgacgagggccagtccgtggtgctgcacgtgcacatcaagcgccacgccatgaaggacctgcccgactccgacgacgccgtggcccagtggtgccgcgacaagacgtggagaaggacgccctgctggacaagcacaacgccgaggacaccactccggccaggaggtgcaccacgtgggccgccccatcaagtccctgctggtggtgatctcctggatggtggtgatcatcttcggcgccctgaagacctgcagtggtcctccctgctgtcctcctggaagggcaaggccactccgccatcggcctgggcatcgccaccctgctgatgcacgtgctggtggtgactcccaggccgaccgctccaaccccgccaaggtgccccccgccaagctgaacaccgagctgtcctcctccaagaaggtgaccaacaaggagaacTGA ctcgagSEQ ID NO: 109 CprocLPAAT2 ggtaccATGgccatccccgccgccgtggccgtgatccccatcggcctgctgacatcatctccggcctgatcgtgaacctgatccaggccgtggtgtacgtgctgatccgccccctgtccaagaacctgtaccgcaagatcaacaagcccatcgccgagctgctgtggctggagctgatctggctggtggactggtgggccggcatcaaggtggaggtgtacgccgactccgagaccctggagtccatgggcaaggagcacgccctgctgatctgcaaccaccgctccgacatcgactggctggtgggctgggtgctggcccagcgcgcccgctgcctgggctccgccctggccatcatgaagaagtccgccaagttcctgcccgtgatcggctggtccatgtggttctccgactacatcttcctggaccgcacctgggagaaggacgagaagaccctgaagtccggcttcgagcgcctggccgacttccccatgcccttctggctggccctgttcgtggagggcacccgcttcaccaaggccaagctgctggccgcccaggagttcgccgcctcccgcggcctgcccgtgccccagaacgtgctgatcccccgcaccaagggcttcgtgaccgccgtgacccacatgcgctcctacgtgcccgccatctacgactgcaccgtggacatctccaaggcccaccccgccccctccatcctgcgcctgatccgcggccagtcctccgtggtgaaggtgcagatcacccgccactccatgcaggagctgcccgagacccccgacggcatctcccagtggtgcatggacctgttcgtgaccaaggacgccttcctggagaagtaccactccaaggacatcttcggctccctgcccgtgcacgacatcggccgccccgtgaagtccctgatcgtggtgctgtgctggtactccctgatggccttcggcactacaagttcttcatgtggtcctccctgctgtcctcctgggagggcatcctgtccctgggcctggtgctgatcgtgatcgccatcgtgatgcagatcctgatccagtcctccgagtccgagcgctccacccccgtgaagtccgtgcagaaggacccctccaaggagaccctgctgcagaacTGA ctcgag SEQ ID NO: 110 CavigGPAT9ggtaccATGgccaccggcggctccctgaagccctcctcctccgacctggacctggaccaccccaacatcgaggactacctgccctccggctcctccatcaacgagcccgccggcaagctgcgcctgcgcgacctgctggacatctcccccaccctgaccgaggccgccggcgccatcgtggacgactccttcacccgctgatcaagtccatcccccgcgagccctggaactggaacctgtacctgttccccctgtggtgcatcggcgtgctgatccgctacttcatcctgttccccggccgcgtgatcgtgctgaccatgggctggatcaccgtgatctcctcatcatcgccgtgcgcgtgctgctgaagggccacgacgccctgcagatcaagctggagcgcctgatcgtgcagctgctgtgctcctcatcgtggcctcctggaccggcgtggtgaagtaccacggcccccgcccctccatccgccccaagcaggtgtacgtggccaaccacacctccatgatcgacttcttcatcctggaccagatgaccgtgttctccgtgatcatgcagaagcaccccggctgggtgggcctgctgcagtccaccctgctggagtccgtgggctgcatctggacgaccgcgccgaggccaaggaccgcggcatcgtggccaagaagctgtgggaccacgtgcacggcgagggcaacaaccccctgctgatcttccccgagggcacctgcgtgaacaacaactactccgtgatgttcaagaagggcgccttcgagctgggctgcaccgtgtgccccgtggccatcaagtacaacaagatcttcgtggacgccttctggaactccaagaagcagtccttcacccgccacctgctgcagctgatgacctcctgggccgtggtgtgcgacgtgtggtacttggagccccagaccctgaagcccggcgagacccccatcgagttcgccgagcgcgtgcgcgacatcatctccgcccgcgccggcctgaagaaggtgccctgggacggctacctgaagtactcccgcccctcccccaagcaccgcgagcgcaagcagcagaccttcgccgagtccgtgctgcagcgcctggaggagTGA ctcgag SEQ ID NO: 111 ChookGPAT9-1 ggtaccATGgccaccgccggctccctgaagccctcccgctccgagctggacttcgaccgccccaacatcgaggactacctgccctccggctcctccatcatcgagcccgccggcaagctgcgcctgcgcgacctgctggacatctcccccaccctgaccgaggccgccggcgccatcgtggacgactccttcacccgctgatcaagtccaacccccccgagccctggaactggaacatctacctgttccccctgtggtgcttcggcgtgctgatccgctacctgatcctgttccccgcccgcgtgatcgtgctgaccatcggctggatcatcttcctgtcctccttcatccccgtgcacctgctgctgaagggccacgacgccctgcgcatcaagctggagcgcctgctggtggagctgatctgctcatcttcgtggcctcctggaccggcgtggtgaagtaccacggcccccgcccctccatccgccccaagcaggtgtacgtggccaaccacacctccatgatcgacttcttcatcctggaccagatgaccgtgttctccgtgatcatgcagaagcaccccggctgggtgggcctgctgcagtccaccctgctggagtccgtgggctgcatctggttcgaccgcgccgaggccaaggaccgcggcatcgtggccaagaagctgtgggaccacgtgcacggcgagggcaacaaccccctgctgatcttccccgagggcacctgcgtgaacaacaactactccgtgatgttcaagaagggcgccttcgagctgggctgcaccgtgtgccccgtggccatcaagtacaacaagatcttcgtggacgccttctggaactccaagaagcagtccttcacccgccacctgctgcagctgatgacctcctgggccgtggtgtgcgacgtgtggtacttggagccccagaccctgaagcccggcgagacccccatcgagttcgccgagcgcgtgcgcgacatcatctccgtgcgcgccggcctgaagaaggtgccctgggacggctacctgaagtactcccgcccctcccccaagcacaccgagcgcaagcagcagaacttcgccgagtccgtgctgcagcgcctggagaagaagTGA ctcgag SEQ ID NO: 112 CignGPAT9-1 ggtaccATGgccaccggcggccgcctgaagccctcctcctccgagctggacctggaccgcgccaacaccgaggactacctgccctccggctcctccatcaacgagcccgtgggcaagctgcgcctgcgcgacctgctggacatctcccccaccctgaccgaggccgccggcgccatcgtggacgactccttcacccgctgcttcaagtccatcccccccgagccctggaactggaacatctacctgttccccctgtggtgatcggcgtgctgatccgctacttcatcctgttccccgcccgcgtgatcgtgctgaccatcggctggatcaccgtgatctcctcatcaccgccgtgcgcacctgctgaagggccacaacgccctgcagatcaagctggagcgcctgatcgtgcagctgctgtgctcctcatcgtggcctcctggaccggcgtggtgaagtaccacggcccccgcccctccatccgccccaagcaggtgtacgtggccaaccacacctccatgatcgacacctgatcctggaccagatgaccgtgactccgtgatcatgcagaagcaccccggctgggtgggcctgctgcagtccaccctgctggagtccgtgggctgcatctggacaaccgcgccgaggccaaggaccgcgagatcgtggccaagaagctgtgggaccacgtgcacggcgagggcaacaaccccctgctgatcaccccgagggcacctgcgtgaacaaccactactccgtgatgacaagaagggcgccacgagctgggctgcaccgtgtgccccgtggccatcaagtacaacaagatcacgtggacgccactggaactcccgcaagcagtccacaccatgcacctgctgcagctgatgacctcctgggccgtggtgtgcgacgtgtggtacaggagccccagaccctgaagcccggcgagaccgccatcgagacgccgagcgcgtgcgcgacatcatctccgtgcgcgccggcctgaagaaggtgccctgggacggctacctgaagtactcccgcccctcccccaagcaccgcgagtccaagcagcagtccacgccgagtccgtgctgcgccgcctggaggagaagTGA ctcgag SEQ ID NO: 113 CignGPAT9-2 ggtaccATGgccaccggcggccgcctgaagccctcctcctccgagctggacctggaccgcgccaacaccgaggactacctgccctccggctcctccatcaacgagcccgtgggcaagctgcgcctgcgcgacctgctggacatctcccccaccctgaccgaggccgccggcgccatcgtggacgactccacacccgctgatcaagtccatcccccccgagccctggaactggaacatctacctgaccccctgtggtgatcggcgtgctgatccgctacttcatcctgaccccgcccgcgtgatcgtgctgaccatcggctggatcaccgtgatctcctcatcaccgccgtgcgcacctgctgaagggccacaacgccctgcagatcaagctggagcgcctgatcgtgcagctgctgtgctcctccacgtggcctcctggaccggcgtggtgaagtaccacggcccccgcccctccatccgccccaagcaggtgtacgtggccaaccacacctccatgatcgacacctgatcctggaccagatgaccgtgactccgtgatcatgcagaagcaccccggctgggtgggcctgctgcagtccaccctgctggagtccgtgggctgcatctggacaaccgcgccgaggccaaggaccgcgagatcgtggccaagaagctgtgggaccacgtgcacggcgagggcaacaaccccctgctgatcaccccgagggcacctgcgtgaacaaccactactccgtgatgacaagaagggcgccacgagctgggctgcaccgtgtgccccgtggccatcaagtacaacaagatcacgtggacgccactggaactccaagaagcactccacacccgccacctgctgcagctgatgacctcctgggccgtggtgtgcgacgtgtggtacaggagccccagaccctgaagcccggcgagacccccatcgagacgccgagcgcgtgcgcgacatcatctccgtgcgcgccgacctgaagaaggtgccctgggacggctacctgaagtactcccgcccctcccccaagcaccgcgagcgcaagcagcagaagacgccgagtccgtgctgcgccgcctggaggagaagTGA ctcgag SEQ ID NO: 114 CpalGPAT9-1 ggtaccATGgccaccgccggccgcctgaagccctcctcctccgagctggagctggacctggaccgccccaacatcgaggactacctgccctccggctcctccatcaacgagcccgccggcaagctgcgcctgcgcgacctgctggacatctcccccatgctgaccgaggccgccggcgccatcgtggacgactccacacccgctgatcaagtccatcccccccgagccctggaactggaacatctacctgttccccctgtggtgatcggcgtgctgatccgctacctgatcctgaccccgcccgcgtgatcgtgctgaccgtgggctggatcaccgtgatctcctccacatcaccgtgcgcacctgctgaagggccacgactccctgcgcatcaagctggagcgcctgatcgtgcagctgttctgctcctccacgtggcctcctggaccggcgtggtgaagtaccacggcccccgcccctccatccgcccccagcaggtgtacgtggccaaccacacctccatgatcgacttcatcatcctgaaccagatgaccgtgactccgccatcatgcagaagcaccccggctgggtgggcctgatccagtccaccatcctggagtccgtgggctgcatctggacaaccgcgccgaggccaaggaccgcgagatcgtggccaagaagctgctggaccacgtgcacggcgagggcaacaaccccctgctgatcaccccgagggcacctgcgtgaacaaccactactccgtgatgacaagaagggcgccacgagctgggctgcaccgtgtgccccgtggccatcaagtacaacaagatcacgtggacgccttctggaactccaagaagcagtccacaccatgcacctgctgcagctgatgacctcctgggccgtggtgtgcgacgtgtggtacaggagccccagaccctgaagcccggcgagacccccatcgagacgccgagcgcgtgcgcgacatcatctccgtgcgcgccggcctgaagaaggtgccctgggacggctacctgaagtactcccgcccctcccccaagcaccgcgagcgcaagcagcagtccacgccgagtccgtgctgcgccgcctggagaagcgcTGA ctcgag SEQ ID NO: 115 CpalGPATt9-2ggtaccATGgccaccgccggccgcctgaagccctcctcctccgagctggagctggacctggaccgccccaacatcgaggactacctgccctccggctcctccatcaacgagcccgccggcaagctgcgcctgcgcgacctgctggacatctcccccatgctgaccgaggccgccggcgccatcgtggacgactccacacccgctgatcaagtccatcccccccgagccctggaactggaacatctacctgttccccctgtggtgatcggcgtgctgatccgctacctgatcctgaccccgcccgcgtgatcgtgctgaccgtgggctggatcaccgtgatctcctccacatcaccgtgcgcacctgctgaagggccacgactccctgcgcatcaagctggagcgcctgatcgtgcagctgttctgctcctccacgtggcctcctggaccggcgtggtgaagtaccacggcccccgcccctccatccgcccccagcaggtgtacgtggccaaccacacctccatgatcgacttcatcatcctgaaccagatgaccgtgactccgccatcatgcagaagcaccccggctgggtgggcctgatccagtccaccatcctggagtccgtgggctgcatctggacaaccgcgccgaggccaaggaccgcgagatcgtggccaagaagctgctggaccacgtgcacggcgagggcaacaaccccctgctgatcaccccgagggcacctgcgtgaacaaccactactccgtgatgttcaagaagggcgccttcgagctgggctgcaccgtgtgccccgtggccatcaagtacaacaagatcttcgtggacgccttctggaactccaagaagctgtcatcaccatgcacctgctgcagctgatgacctcctgggccgtggtgtgcgacgtgtggtacttggagccccagaccctgaagcccggcgagacccccatcgagttcgccgagcgcgtgcgcgacatcatctccgtgcgcgccggcctgaagaaggtgccctgggacggctacctgaagtactcccgcccctcccccaagcaccgcgagcgcaagcagcagaccttcgccgagtccgtgctgcgccgcctggaggagaagggcaacgtggtgcccaccgtgaacTGA ctcgagSEQ ID NO: 116 CavigDGAT1 ggtaccATGgccatcgccgacggcggcatcatcggcgccgccggctccatctccgccctgaccgccgacaccgaccccccctccctgcgccgccgcaacgtgcccgccggccaggcctccgccgtgtccgccttctccaccgagtccatggccaagcacctgtgcgacccctcccgcgagccctccccctcccccaagtcctccgacgacggcaaggaccccgacatcggctccgtggactccctgaacgagaagccctcctcccccgccgccggcaagggccgcctgcagcacgacctgcgatcacctaccgcgcctcctcccccgcccaccgcaaggtgaaggagtcccccctgtcctcctccaacatcttcaagcagtcccacgccggcctgttcaacctgtgcgtggtggtgctggtggccgtgaactcccgcctgatcatcgagaacctgatgaagtacggcctgctgatcaagaccggcttctggttctcctcccgctccctgcgcgactggcccctgttcatgtgctgcctgtccctgcccatcttccccctggccgccttcctggtggagaagctggcccagaagaaccgcctgcaggagcccaccgtggtgtgctgccacgtgctgatcacctccgtgtccatcctgtaccccgtgctggtgatcctgcgctgcgactccgccgtgctgtccggcgtggccctgatgctgttcgcctgcatcgtgtggctgaagctggtgtcctacgcccactccaactacgacatgcgctacgtggccaagtccctggacaagggcgagcccgtggtggactccgtgatcgccgaccacccctaccgcgtggactacaaggacctggtgtacttcatggtggcccccaccctgtgctaccagctgtcctaccccctgaccccctgcgtgcgcaagtcctggatcgcccgccaggtgatgaagctggtgctgttcaccggcgtgatgggcttcatcgtggagcagtacatcaaccccatcgtgcagaactccaagcaccccctgaagggcgacctgctgtacgccatcgagcgcgtgctgaagctgtccgtgcccaacctgtacgtgtggctgtgcatgttctactgcttcttccacctgtggctgaacatcctggccgagctgatctgcttcggcgaccgcgagttctacaaggactggtggaacgccaagaccgtggaggagtactggcgcatgtggaacatgcccgtgcacaagtggatggtgcgccacatctacttcccctgcctgcgcaacggcatcccccgcggcgtggccgtgctgatcgccttcctggtgtccgccgtgttccacgagctgtgcatcgccgtgccctgccacgtgttcaagctgtgggccttcatcggcatcatgttccaggtgcccctggtgctggtgtccaactgcctgcagaagaagttccagtcctccatggccggcaacatgttatctggttcatcttctgcatcttcggccagcccatgtgcgtgctgctgtactaccacgacctgatgaaccgcaagggctcccgcatcgacTGA ctcgag SEQ ID NO: 117 ChookDGAT1-1ggtaccATGgccatcgccgacggcggctccgccggcgccgccggctccatctccggctccgacccctccccctccaccgccccctccctgcgccgccgcaacgcctccgccggccaggccttctccaccgagtccatggcccgcgacctgtgcgacccctcccgcgagccctccctgtcccccaagtcctccgacgacggcaaggaccccgccgacgacatcggcgccgccgactccgtggactccggcggcgtgaaggacgagaagccctcctcccaggccgccgccaaggcccgcctggagcacgacctgcgatcacctaccgcgcctcctcccccgcccaccgcaaggtgaaggagtcccccctgtcctcctccaacatcttcaagcagtcccacgccggcctgttcaacctgtgcgtggtggtgctggtggccgtgaactcccgcctgatcatcgagaacctgatgaagtacggcctgctgatcaagaccggcttctggttctcctcccgctccctgcgcgactggcccctgttcatgtgctgcctgtccctgcccatcaccccctggccgccttcctggtggagaagctggcccagaagaaccgcctgcaggagcccaccgtggtgtgctgccacgtgatcatcacctccgtgtccatcctgtaccccgtgctggtgatcctgcgctgcgactccgccgtgctgtccggcgtggccctgatgctgttcgcctgcatcgtgtggctgaagctggtgtcctacgcccacgccaactacgacatgcgctccgtggccaagtccctggacaagggcgagaccgtggccgactccgtgatcgtggaccacccctaccgcgtggactacaaggacctggtgtacttcatggtggcccccaccctgtgctaccagctgtcctaccccctgaccccctacgtgcgcaagtcctgggtggcccgccaggtgatgaagctggtgctgttcaccggcgtgatgggcttcatcgtggagcagtacatcaaccccatcgtgcagaactccaagcaccccctgaagggcgacctgctgtacgccatcgagcgcgtgctgaagctgtccgtgcccaacctgtacgtgtggctgtgcatgttctactgcttcttccacctgtggctgaacatcctggccgagctgacctgcttcggcgaccgcgagttctacaaggactggtggaacgccaagaccgtggaggagtactggcgcatgtggaacatgcccgtgcacaagtggatggtgcgccacatctacttcccctgcctgcgcaacggcatcccccgcggcgtggccgtgctgatcgccttcctggtgtccgccgtgttccacgagctgtgcatcgccgtgccctgccacgtgttcaagctgtgggccttcatcggcatcatgttccaggtgcccctggtgctggtgtccaactgcctgcagaagaagttccagtcctccatggccggcaacatgttatctggttcatcttctgcatcttcggccagcccatgtgcgtgctgctgtactaccacgacctgatgaaccgcaagggctcccgcatcgacTGA ctcgagSEQ ID NO: 118 CavigLPCAT ggtaccATGggcctggtgtccgtggccgccgccatcggcgtgtccgtgcccgtggcccgcttcctgctgtgcttcctggccaccatccccgtgtccttcctgtggcgcctggtgcccggccgcctgcccaagcacctgtactccgccgcctccggcgccatcctgtcctacctgtcatcggcgcctcctccaacctgcacttcatcgtgcccatgaccctgggctacctgtccatgctgttcttccgcccatctccggcctgctgaccttcttcctgggcttcggctacctgatcggctgccacgtgtactacatgtccggcgacgcctggaaggagggcggcatcgacgccaccggcgccctgatggtgctgaccctgaaggtgatctcctgctccatgaactacaacgacggcctgctgaaggaggagggcctgcgcgagtcccagaagaagaaccgcctgaccaagatgccctccctgatcgagtacttcggctactgcctgtgctgcggctcccacttcgccggccccgtgtacgagatgaaggactacctggagtggaccgagggcaagggcatctggtcccgctcccagaaggagcccaagccctccccatcggcggcgccctgcgcgccatcatccaggccgccgtgtgcatggccatgtacctgtacctggtgccccaccaccccctgacccgcttcaccgagcccgtgtactacgagtggggatcttccgccgcctgtcctaccagtacatggccgccctgaccgcccgctggaagtactacttcatctggtccatctccgaggcctccctgatcatctccggcctgggcttctccggctggaccgagtcctccccccccaagccccgctgggaccgcgccaagaacgtggacatcatcggcgtggagttcgccaagtcctccgtgcagctgcccctggtgtggaacatccaggtgtccatctggctgcgccactacgtgtacgaccgcctggtgcagaacggcaagcgccccggcttcaccagctgctggccacccagaccgtgtccgccgtgtggcacggcctgtaccccggctacatcatatatcgtgcagtccgccctgatgatcgccggctcccgcgtgatctaccgctggcagcaggccgtgccccccaagatgggcctggtgaagaacatcttcgtgttcttcaacttcgcctacaccctgctggtgctgaactactccgccgtgggcttcatggtgctgtccatgcacgagaccctggcctcctacggctccgtgtactacatcggcaccatcctgcccatcaccctgatcctgctgtcctacgtgatcaagcccggcaagcccgcccgctccaaggcccacaaggagcagTGA ctcgag SEQ ID NO: 119 CpalLPCAT ggtaccATGgagctgggctccgtggccgccgccatcggcgtgtccgtgcccgtggcccgcttcctgctgtgcttcctggccaccatccccgtgtccttcctgtggcgcctggtgcccggccgcctgcccaagcacctgtactccgccgcctccggcgccatcctgtcctacctgtcatcggcccctcctccaacctgcacttcatcgtgcccatgaccctgggctacctgtccatgctgttcttccgcccatctccggcctgctgaccttcttcctgggcttcggctacctgatcggctgccacgtgtactacatgtccggcgacgcctggaaggagggcggcatcgacgccaccggcgccctgatggtgctgaccctgaaggtgatctcctgctccatcaactacaacgacggcctgctgaaggaggagggcctgcgcgagtcccagaagaagaaccgcctgaccaagatgccctccctgatcgagtacatcggctactgcctgtgctgcggctcccacttcgccggccccgtgtacgagatgaaggactacctggagtggaccgagggcaagggcgtgtggtcccactccgagaaggagcccaagccctccccatcggcggcgccctgcgcgccatcatccaggccgccgtgtgcatggccatgtacatgtacctggtgccccaccaccccctgtcccgatcaccgagcccgtgtactacgagtggggcacttccgccgcctgtcctaccagtacatggccggcctgaccgcccgctggaagtactacttcatctggtccatctccgaggcctccctgatcatctccggcctgggcttctccggctggaccgagtcctccccccccaagccccgctgggaccgcgccaagaacgtggacatcatcggcgtggagttcgccaagtcctccgtgcagctgcccctggtgtggaacatccaggtgtccacctggctgcgccactacgtgtacgaccgcctggtgcagaacggcaagcgccccggcttcaccagctgctggccacccagaccgtgtccgccatctggcacggcctgtaccccggctacatcatatatcgtgcagtccgccctgatgatcgccggctcccgcgtgatctaccgctggcagcaggccgtgccccccaagatgggcctggtgaagaacatcttcgtgttcttcaacttcgcctacaccctgctggtgctgaactactccgccgtgggcttcatggtgctgtccatgcacgagaccctggcctcctacggctccgtgtactacatcggcaccatcctgcccatcaccctgatcctgctgtcctacgtgatcaagcccggcaagcccgcccgctccaaggcccacaaggagcagTGA ctcgag SEQ ID NO: 120 CpauLPCAT ggtaccATGgagctggagatcggctccgtggccgccgccatcggcgtgtccgtgcccgtggcccgatcctgctgtgatcaggccaccatccccgtgtccttcctgtgccgcctgctgcccgcccgcctgcccaagcacctgtactccgccgcctccggcgccatcctgtcctacctgtcatcggcccctcctccaacctgcacttcatcgtgcccatgtccctgggctacctgtccatgctgttcttccgccccttctccggcctgctgaccttcttcagggatcggctacctgatcggctgccacgtgtactacatgtccggcgacgcctggaaggagggcggcatcgacgccaccggcgccctgatggtgctgaccctgaaggtgatctcctgctccatcaactacaacgacggcctgctgaaggaggagggcctgcgcgagtcccagaagaagaaccgcctgaccaagatgccctccctgatcgagtacttcggctactgcctgtgctgcggctcccacttcgccggccccgtgtacgagatgaaggactacctggagtggaccgagggcaagggcatctggtcccgctccgagaaggaccccaagccctccccatcggcggcgccctgcgcgccatcatccaggccgccgtgtgcatggccatgcacatgtacctggtgccccaccaccccctgacccgcttcaccgagcccgtgtactacgagtggggatcttccgccgcctgtcctaccagtacatggccgcccagaccgcccgctggaagtactacttcatctggtccatctccgaggcctccctgatcatctccggcctgggcttctccggctggaccgagtcctccccccccaagccccgctgggacaaggccaagaacgtggacatcatcggcgtggagttcgccaagtcctccgtgcagctgcccctggtgtggaacatccaggtgtccacctggctgcgccactacgtgtacgaccgcctggtgcagaacggcaagcgccccggcttcttccagctgctggccacccagaccgtgtccgccgtgtggcacggcctgtaccccggctacatcatcttcttcgtgcagtccgccctgatgatcgccggctcccgcgtgatctaccgctggcagcaggccgtgccccagaagatgggcctggtgaagaacatcttcgtgttcttcaacttcgcctacaccctgctggtgctgaactactccgccgtgggcttcatggtgctgtccatgcacgagaccctggcctcctacggctccgtgtactacatcggcaccatcctgcccatcaccctgatcctgctgtcctacgtgatcaagcccggcaagcccacccgctccaaggtgcacaaggagcagTGA ctcgag SEQ ID NO: 121 CschuLPCAT ggtaccATGgagctggagatggagcccctggccgccgccatcggcgtgtccgtggccgtgttccgcttcctggtgtgcttcatcgccaccatccccgtgtccttcatctgccgcctggtgcccggcggcctgccccgccacctgttctccgccgcctccggcgccgtgctgtcctacctgtcatcggatctcctccaacctgcacttcctggtgcccatgaccctgggctacctgtccatgatcctgttccgccgatctgcggcatcctgaccttcttcctgggcttcggctacctgatcggctgccacgtgtactacatgtccggcgacgcctggaaggagggcggcatcgacgccaccggcgccctgatggtgctgaccctgaaggtgatctcctgctccatcaactacaacgacggcctgctgaaggaggagggcctgcgcgagtcccagaagaagaaccgcctgatccgcctgccctccctgatcgagtacttcggctactgcctgtgctgcggctcccacttcgccggccccgtgtacgagatgaaggactacctggactggaccgagggcaagggcatctggtcccactccgagaagggccccaagccctcccccctgcgcgccgccctgcgcgccatcatccaggccggcttctgcatggccatgtacctgtacctggtgccccactaccccctgacccgcttcaccgaccccgtgtactacgagtggggcatcctgcgccgcctgtcctaccagtacatggcctccttcaccgcccgctggaagtactacttcatctggtccatctccgaggcctccctgatcatctccggcctgggcttctccggctggaccgagtcctccccccccaagccccgctgggaccgcgccaagaacgtggacatcctgggcgtggagctggccaagtcctccgtgcagatccccctggtgtggaacatccaggtgtccacctggctgcgccactacgtgtacgaccgcctggtgcagaacggcaagcgccccggatcctgcagctgctggccacccagaccgtgtccgccatctggcacggcgtgtaccccggctacctgatcacttcgtgcagtccgccctgatgatcgccggctcccgcgccatctaccgctggcagcaggccgtgccccccaagatgtccctggtgaagaacaccctggtgttcttcaacttcgcctacaccctgctggtgctgaactactccgccgtgggcttcatggtgctgtccatgcacgagaccctggcctcctacggctccgtgtactacgtgggcaccatcctgcccgtgaccctgatcctgctgggctacgtgatcaagcccggcaagtccccccgctccaaggcctccaaggagcagTGA ctcgag SEQ ID NO: 122 CavigPLA2-1 ggtaccATGaacttcgacttcctgtccaacatcccctggttcggcgccaaggcctccgacaacgccggctcctcatcggctccgccaccatcgtgatccagcagcccccccccgtgtcccgcggcttcgacatccgccactggggctggccctggtccgtgctgtccgtgctgccctggggcaagcccggctgcgacgagctgcgcgccccccccaccaccatcaaccgccgcctgaagcgcaacgccacctccatgcactcctccgccgtgcgcggcaacgccgaggccgcccgcgtgcgcttccgcccctacgtgtccaaggtgccctggcacaccggcttccgcggcctgctgtcccagctgttcccccgctacggccactactgcggccccaactggtcctccggcaagaacggcggctcccccgtgtgggaccagcgccccatcgactggctggactactgctgctactgccacgacatcggctacgacacccacgaccaggccaagctgctggaggccgacctggccttcctggagtgcctggagcgcccctcctaccccaccaagggcgacgcccacgtggcccacatgtacaagaccatgtgcgtgaccggcctgcgcaacgtgctgatcccctaccgcacccagctgctgcgcctgaactcccgccagcccctgatcgacttcggctggctgtccaacgccgcctggaagggctggaacgcccagaagtccTGActcgag SEQ ID NO: 123 CignPLA2-1 ggtaccATGaacctggacttcctgtccaagatcccctggttcgaggccaaggcctccgagaaccccggcctgaacctgggctccaccaccatcgtgatcaagcagccccgccagggcttcgacatccgccactggggctggccctggtccgtgctgacctggggcaaccgcgtgaccgacgaggtgcacgccccccccaccaccatcaaccgccgcctgaagcgcaacgccaccggccccgccgtgcagggcgacaccgaggccgcccgcctgcgcttccgcccctacgtgtccaaggtgccctggcacaccggcttccgcggcctgctgtcccagctgttcccccgctacggccactactgcggccccaactggtcctccggcaagaacggcggctcccccgtgtgggaccagcgccccatcgactggctggactactgctgctactgccacgacatcggctacgacacccacgaccaggccaagctgctggaggccgacctggccttcctggagtgcctggagcgcccctcctaccccaccaccggcgacgcccacgtggcccacatgtacaagaccatgtgcgtgaccggcctgcgcaacgtgctgatcccctaccgcacccagctgctgcgcctgaacttccgccagcccctgatcgacttcggctggctgtccaacgccgcctggaagggctggtccgcccagaagaccTGA ctcgagSEQ ID NO: 124 CuPSR23PLA2-2 ggtaccATGgtgcacctgccccacaccctgaagctgggcctggtgatcgccatctccatctccggcctgtgcactcctccacccccgcccgcgccctgaacgtgggcatccaggccgccggcgtgaccgtgtccgtgggcaagggctgctcccgcaagtgcgagtccgacactgcaaggtgccccccacctgcgctacggcaagtactgcggcctgatgtactccggctgccccggcgagaagccctgcgacggcctggacgcctgctgcatgaagcacgacgcctgcgtgcaggccaagaacaacgactacctgtcccaggagtgctcccagaacctgctgaactgcatggcctccaccgcatgtccggcggcaagcagacaagggctccacctgccaggtggacgaggtggtggacgtgctgaccgtggtgatggaggccgccctgctggccggccgctacctgcacaagcccTGA ctcgagSEQ ID NO: 125 CprocPLA2-2 ggtaccATGgtgcacctgccccacaccctgaagctgggcctggtgatcgccatctccatctccggcctgtgcctgtcctccacccccgcccgcgccctgaacgtgggcatccaggccgccggcgtgaccgtgtccgtgggcaagggctgctcccgcaagtgcgagtccgacactgcaaggtgccccccacctgcgctacggcaagtactgcggcctgatgtactccggctgccccggcgagaagccctgcgacggcctggacgcctgctgcatgaagcacgacgcctgcgtgcaggccaagaacgacgactacctgtcccaggagtgctcccagaacctgctgaactgcatggcctccaccgcatgtccggcggcaagcagacaagggctccacctgccaggtggacgaggtggtggacgtgctgaccgtggtgatggaggccgccctgctggccggccgctacctgcacaagcccTGA ctcgag

The constructs containing the codon optimized genes described abovedriven by the UTEX 1453 SAD2 promoter, were transformed into strainS7858 or S8714. Transformations, cell culture, lipid production andfatty acid analysis were all carried out as described herein. Thetransgenic strains were selected for their ability to grow on melibiose.Stable transformants were grown under standard lipid productionconditions at pH5 (for transgenic strains generated in the strain S7858)or at pH7 (for the transgenic strains generated in the strain S8174) forfatty acid analysis.

Expression of LPAATs

In WO2013/158938 we disclosed that Cocos nucifera LPAAT enzymes exhibitchain length specificity for the fatty acid acyl-CoA that it attach tothe glycerol backbone. We disclosed the impact of expressing CnLPAAT ina transgenic strain also expressing a laurate specific thioesterase. Inthis example we transformed 5 LPAAT enzymes derived from C8-C10 richCuphea species and the CnLPAAT into S7858, and the remaining 8 LPAATenzymes were transformed into S8174. The resulting fatty acid profilesfrom a set of representative transgenic lines arising from thesetransformations are shown in Tables 16 and 17. Expression of these genesas shown in Table 16 resulted in increases in C8:0 and/or- C10:0 fattyacid accumulation.

TABLE 16 Fatty acid profiles of representative transgenic strains ofS7858 expressing optimized versions of the CpauLPAAT1, CpalLPAAT1,CignLPAAT1, CprocLPAAT1, ChookLPAAT1 and CnLPAAT1. Sample ID C8:0 C10:0C12:0 C8-C10 S6165  0.00  0.00 0.05  0.00 S7858 11.70 23.36 0.48 35.06CpauLPAAT1 @ SAD2-1vD locus CprocLPAAT1 @ SAD2-1vD locus Sample ID C8:0C10:0 C12:0 C8-C10 Sample ID C8:0 C10:0 C12:0 C8-C10 S7858; D4289-7 12.69 25.06 0.51 37.75 S7858; D4292-15 11.86 24.05 0.46 35.91 S7858;D4289-12 11.98 24.54 0.48 36.52 S7858; D4292-11 11.49 24.01 0.48 35.50S7858; D4289-2  11.68 24.14 0.49 35.82 S7858; D4292-22 11.49 23.81 0.4735.30 S7858; D4289-13 11.53 24.18 0.49 35.71 S7858; D4292-3  11.46 23.760.46 35.22 S7858; D4289-11 11.47 23.85 0.46 35.32 S7858; D4292-24 11.3823.64 0.46 35.02 CpaiLPAAT1 @ SAD2-1vD locus ChookLPAAT1 @ SAD2-1vDlocus Sample ID C8:0 C10:0 C12:0 C8-C10 Sample ID C8:0 C10:0 C12:0C8-C10 S7858; D4290-3  13.43 25.04 0.52 38.47 S7858; D4293-4  11.0924.48 0.51 35.57 S7858; D4290-25 12.98 24.75 0.51 37.73 S7858; D4293-1612.03 24.24 0.48 36.27 S7858; D4290-5  12.27 25.00 0.52 37.27 S7858;D4293-6  11.83 23.79 0.48 35.62 S7858; D4290-12 11.98 24.21 0.48 36.19S7858; D4293-2  11.81 23.69 0.47 35.50 S7858; D4290-22 11.91 23.86 0.4935.77 S7858; D4293-12 11.65 23.11 0.49 34.76 CignLPAAT1 @ SAD2-1vD locuCnLPAAT1 @ SAD2-1vD locus Sample ID C8:0 C10:0 C12:0 C8-C10 Sample IDC8:0 C10:0 C12:0 C8-C10 S7858; D4291-13 12.95 24.78 0.52 37.73 S7858;D4404-11 12.30 24.31 0.47 36.61 S7858; D4291-20 12.13 24.63 0.49 36.76S7858; D4404-6  12.03 24.02 0.46 36.05 S7858; D4291-15 12.12 24.35 0.4736.47 S7858; D4404-13 11.48 23.98 0.46 35.46 S7858; D4291-22 11.94 24.500.47 36.44 S7858; D4404-2  11.54 23.71 0.46 35.25 S7858; D4291-7  12.1123.14 0.50 35.25 S7858; D4404-1  11.76 23.36 0.48 35.12

TABLE 17 Fatty acid profiles of representative transgenic strains ofS8174 expressing CavigLPAAT1, CavigLPAAT2, CpalLPAAT1, CuPSR23LPAAT1,CkoeLPAAT1, CkoeLPAAT2, CprocLPAAT1 and CprocLPAAT2 before lipasetreatment. Sample ID C8:0 C10:0 C12:0 C8-C10 S7485  0.00 0.00 0.07  0.00S8174 24.32 9.24 0.37 33.56 CavigLPAAT1 @ SAD2-1vD locus CkoeLPAAT1 @SAD2-1vD locus C8- C8- Sample ID C8:0 C10:0 C12:0 C10 Sample ID C8:0C10:0 C12:0 C10 S8174: D4517-23 25.42 9.63 0.39 35.05 S8174; D4728-8 25.44 10.31 0.46 35.75 S8174: D4517-9  25.44 9.61 0.39 35.05 S8174;D4728-10 24.15 9.51 0.43 33.66 S8174: D4517-8  25.09 9.84 0.39 34.93S8174; D4728-5  23.88 9.56 0.45 33.44 S8174: D4517-18 25.20 9.65 0.3934.85 S8174; D4728-6  23.58 9.28 0.40 32.86 S8174: D4517-2  25.20 9.570.37 34.77 S8174; D4728-9  23.47 9.25 0.40 32.72 Cavig LPAAT2 @ SAD2-1vDlocus CkoeLPAAT2-1 @ SAD2-1vD locus C8- C8- Sample ID C8:0 C10:0 C12:0C10 Sample ID C8:0 C10:0 C12:0 C10 S8174: D4518-2  24.25 9.97 0.42 34.22S8174; D4729-2 25.20 9.81 0.43 35.01 S8174: D4518-45 24.09 9.65 0.3933.74 S8174; D4729-1 23.49 10.60 0.46 34.09 S8174: D4518-34 23.94 9.710.38 33.65 S8174; D4729-4 22.25 9.45 0.40 31.70 S8174: D4518-10 24.119.50 0.37 33.61 S8174; D4729-5 18.24 8.22 0.35 26.46 S8174: D4518-4 23.93 9.59 0.39 33.52 CpalLPAAT1 @ SAD2-1vD locus CprocLPAAT2 @ SAD2-1vDlocus C8- C8- Sample ID C8:0 C10:0 C12:0 C10 Sample ID C8:0 C10:0 C12:0C10 S8174: D4519-27 25.06 9.75 0.37 34.81 S8174; D4730-14 24.97 9.920.41 34.89 S8174: D4519-4  23.05 10.74 0.47 33.79 S8174; D4730-13 23.2610.72 0.49 33.98 S8174: D4519-28 24.11 9.54 0.37 33.65 S8174; D4730-1 23.79 10.15 0.49 33.94 S8174: D4519-10 23.57 9.51 0.38 33.08 S8174;D4730-7  23.42 10.13 0.36 33.55 S8174: D4519-12 23.55 9.49 0.38 33.04S8174; D4730-5  23.69 9.49 0.42 33.18 CuPSR23LPAAT2-1 @ SAD2-1vD locusCuPSR23LPAAT4 @ SAD2-1vD locus C8- C8- Sample ID C8:0 C10:0 C12:0 C10Sample ID C8:0 C10:0 C12:0 C10 S8174; D4690-2 25.88 10.62 0.43 36.50S8174; D4731-1 25.94 10.87 0.56 36.81 S8174; D4690-1 24.60 9.82 0.4434.42 S8174; D4731-3 22.79 11.52 0.59 34.31 S8174; D4690-3 24.13 9.620.47 33.75 S8174; D4731-5 22.89 11.22 0.53 34.11 S8174; D4690-4 23.389.97 0.41 33.35 S8174; D4731-2 22.99 11.07 0.45 34.06 S8174; D4731-421.15 9.63 0.43 30.78

To assess the regiospecific activity of novel LPAAT enzymes, oilextracted from some of these transformants were treated with porcinepancreatic lipase, which selectively hydrolyzes the fatty acids at thesn-1 and sn-3 positions from the glycerol unit of the triacylglycerol,leaving monoacylglycerols (MAGs) with fatty acids located only at thesn-2 position. The resulting mixture of monoacylglycrols (2-MAGs), wereisolated by solid phase extraction on an amino propyl cartridge followedby transesterifcation to generate fatty acid methyl esters (FAMEs). Thefatty acid profiles of these FAMEs, which represent the profile of fattyacids at the sn-2 position of the various TAGs, were determined byGC-FID. When compared to the fatty acid profiles fromtransesterification of the oil without lipase treatment, the sn-2 fattyacid profiles show that the expressed LPAAT are selective for the sn-2position.

The sn-2 analyses after lipase treatment disclosed in Table 18 show thatCavigLPAAT1, CpaiLPAAT exhibit selectivity for either C8:0 fatty acidsand CpauLPAAT, CignLPAAT are selective for C10:0 fatty acids,demonstrating that the heterologous LPAATs expressed in these transgenicstrains have activities that acylate at the sn-2 position withpreference for C8:0 or C10:0.

TABLE 18 Fatty acid profiles & sn-2 analysis of representativetransgenic strains of S7858 & S8174 expressing codon optimized versionsof the CnLPAAT1, CpauLPAAT1, CpaiLPAAT1, CignLPAAT1, ChookLPAAT1 andCavigLPAAT1, CavigLPAAT2, CpalLPAAT1  

   

   

   

   

  Fatty Acid FA profile sn-2 FA profile sn-2 FA profile sn-2 FA profilesn-2 FA profile sn-2 C 

   

   

   

   

   

   

   

   

   

   

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  Fatty Acid FA profile sn-2 FA profile sn-2 FA profile sn-2 FA profilesn-2 C 

   

   

   

   

   

   

   

   

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indicates data missing or illegible when filed

Expression of GPATs, DGATs, LPCATs and PLA2s:

The constructs expressing the other acyltransferases (GPAT, DGAT, LPCAT,and PLA2) were transformed into S8174. Stable transformants were grownunder standard lipid production conditions at pH7 and analyzed for fattyacid profiles. Similar to the transgenic lines expressing LPAATs,expression of these genes (GPAT, DGAT, LPCAT, and PLA2) also resulted inincreases in C8:0-C10:0 fatty acid accumulation (Tables 19a, 19b, and20). The data presented shows that we have identified novel GPATs,DGATs, LPCATs and PLA2s that show high specificity for C8-C10 fattyacids. To determine the regiospecificity of the novel GPAT, DGAT, LPCAT,and PLA2 enzymes, sn-2 analysis is performed as disclosed in thisexample and elsewhere herein.

TABLE 19a Fatty acid profiles of representative transgenic strains ofS8174 expressing GPATs and DGATs Sample ID C8:0 C10:0 C12:0 C8-C10 S7485 0.00 0.00 0.07  0.00 S8174 24.61 9.10 0.42 33.71 CavigGPAT9 @ SAD2-1vDlocus CignGPAT9-2 @ SAD2-1vD locus Sample ID C8:0 C10:0 C12:0 C8-C10Sample ID C8:0 C10:0 C12:0 C8-C10 S8174; D4551-8 24.52 9.05 0.36 33.57S8174; D4554-9 24.49 9.13 0.45 33.62 S8174; D4551-7 24.24 9.04 0.3633.28 S8174; D4554-3 24.28 8.90 0.42 33.18 S8174; D4551-2 23.93 8.920.37 32.85 S8174; D4554-7 23.86 8.96 0.43 32.82 S8174; D4551-6 23.638.92 0.41 32.55 S8174; D4554-8 23.99 8.81 0.39 32.80 S8174; D4551-323.35 8.90 0.43 32.25 S8174; D4554-4 23.87 8.78 0.4 32.65 ChookGPAT9-1 @SAD2-1vD locus CpalGPAT9-1 @ SAD2-1vD locus Sample ID C8:0 C10:0 C12:0C8-C10 Sample ID C8:0 C10:0 C12:0 C8-C10 S8174; D4552-6  23.57 9.00 0.3632.57 S8174; D4724-6 25.61 9.52 0.39 35.13 S8174; D4552-4  23.62 8.870.37 32.49 S8174; D4724-7 24.91 9.36 0.41 34.27 S8174; D4552-9  23.398.97 0.40 32.36 S8174; D4724-2 24.43 9.46 0.39 33.89 S8174; D4552-8 23.28 8.80 0.40 32.08 S8174; D4724-5 24.01 9.25 0.39 33.26 S8174;D4552-11 23.18 8.80 0.44 31.98 S8174; D4724-4 24.30 8.93 0.39 33.23CignGPAT9-1 @ SAD2-1vD locus CpalGPAT9-2 @ SAD2-1vD locus Sample ID C8:0C10:0 C12:0 C8-C10 Sample ID C8:0 C10:0 C12:0 C8-C10 S8174; D4553-1225.19 9.42 0.40 34.61 S8174; D4725-5 24.24 10.30 0.48 34.54 S8174;D4685-1  24.33 10.24 0.46 34.57 S8174; D4725-6 24.81 9.29 0.41 34.10S8174; D4553-15 25.11 9.33 0.41 34.44 S8174; D4725-7 24.35 9.51 0.4233.86 S8174; D4553-1  24.56 9.50 0.44 34.06 S8174; D4725-8 24.37 9.390.40 33.76 S8174; D4553-6  24.74 9.16 0.40 33.90 S8174; D4725-9 24.289.29 0.41 33.57

TABLE 19b Fatty acid profiles of representative transgenic strains ofS8174 expressing DGATs Sample ID C8:0 C10:0 C12:0 C8-C10 S7485 0.00 0.000.07 0.00 S8174 24.61 9.10 0.42 33.71 Cavig DGAT1 @ SAD2-1vD locusS8174; D4549-7 24.89 9.28 0.36 34.17 S8174; D4549-6 24.53 9.04 0.4733.57 S8174; D4549-4 23.93 8.99 0.41 32.92 S8174; D4549-1 23.93 8.970.38 32.90 S8174; D4549-3 23.76 8.9 0.36 32.66 Chook DGAT1 @ SAD2-1vDlocus S8174; D4550-1 24.67 9.12 0.41 33.79 S8174; D4550-3 24.64 9.060.42 33.70 S8174; D4682-1 23.72 9.68 0.5 33.40 S8174; D4682-2 23.49 9.660.41 33.15 S8174; D4550-2 22.42 8.81 0.41 31.23

TABLE 20 Fatty acid profiles of representative transgenic strains ofS8174 expressing LPCATs and PLA2s Sample ID C8:0 C10:0 C12:0 C8-C10S7485  0.00 0.00 0.07  0.00 S8174 24.61 9.10 0.42 33.71 Cavig LPCAT @SAD2-1vD locus Cavig PLA2-1 @ SAD2-1vD locus Sample ID C8:0 C10:0 C12:0C8-C10 Sample ID C8:0 C10:0 C12:0 C8-C10 S8174; D4555-1 26.6 9.38 0.4735.98 S8174; D4732-1 26.31 11.24 0.60 37.55 S8174; D4555-3 26.4 9.470.39 35.87 S8174; D4732-2 25.30 11.88 0.50 37.18 S8174; D4688-1 25.959.67 0.44 35.62 S8174; D4732-3 25.29 11.01 0.48 36.30 S8174; D4688-325.47 9.89 0.44 35.36 S8174; D4732-4 25.30 11.00 0.47 36.30 S8174;D4555-2 25.52 9.55 0.36 35.07 S8174; D4732-5 25.07 11.20 0.44 36.27 CpauLPCAT @ SAD2-1vD locus CignPLA2-1 @ SAD2-1vD locus Sample ID C8:0 C10:0C12:0 C8-C10 Sample ID C8:0 C10:0 C12:0 C8-C10 S8174; D4556-3 25.55 9.210.43 34.76 S8174; D4734-6 26.39 11.34 0.47 37.73 S8174; D4556-4 25.249.46 0.41 34.70 S8174; D4734-1 26.17 10.90 0.46 37.07 S8174; D4689-724.63 9.86 0.43 34.49 S8174; D4734-5 25.58 11.12 0.57 36.70 S8174;D4556-1 25.18 9.13 0.42 34.31 S8174; D4734-4 25.48 11.17 0.57 36.65S8174; D4689-6 24.05 9.89 0.48 33.94 S8174; D4734-2 24.75 11.32 0.4636.07 Cpal LPCAT @ SAD2-1vD locus CuPSR23PLA2-2 @ SAD2-1vD locus SampleID C8:0 C10:0 C12:0 C8-C10 Sample ID C8:0 C10:0 C12:0 C8-C10 S8174;D4726-4 26.34 9.76 0.41 36.10 S8174; D4735-5 25.81 11.16 0.44 36.97S8174; D4726-2 25.92 9.9 0.44 35.82 S8174; D4735-1 25.95 10.92 0.4736.87 S8174; D4726-3 26.15 9.62 0.41 35.77 S8174; D4735-8 25.54 10.910.42 36.45 S8174; D4726-5 26.09 9.55 0.41 35.64 S8174; D4735-7 25.4510.95 0.44 36.40 S8174; D4726-1 25.64 9.57 0.39 35.21 S8174; D4735-625.51 10.88 0.41 36.39 Cschu LPCAT @ SAD2-1vD locus Cproc PLA2-2 @SAD2-1vD locus Sample ID C8:0 C10:0 C12:0 C8-C10 Sample ID C8:0 C10:0C12:0 C8-C10 S8174; D4727-1  26.24 9.95 0.45 36.19 S8174; D4736-2 25.6010.87 0.42 36.47 S8174; D4727-7  26.26 9.84 0.42 36.10 S8174; D4736-425.55 10.76 0.40 36.31 S8174; D4727-9  26.13 9.87 0.42 36.00 S8174;D4736-3 25.40 10.87 0.36 36.27 S8174; D4727-11 25.99 9.97 0.44 35.96S8174; D4736-5 25.45 10.46 0.39 35.91 S8174; D4727-16 26.28 9.68 0.4435.96 S8174; D4736-1 24.34 11.06 0.48 35.40

Example 7: Expression of LPAAT and/or DGAT in Prototheca to Produce HighSOS and Low Trisaturated TAGs

In this example we describe genetically engineered Prototheca moriformisstrains in which we have modified fatty acid and triacylglycerolbiosynthesis to maximize the accumulation of Stearoyl-Oleoyl-Stearoyl(SOS) TAGs, and minimize the production of trisaturated TAGs. Tailoredoils from these strains resemble plant seed oils known as “structuringfats”, which have high proportions of Saturated-Oleate-Saturated TAGsand low levels of trisaturates. These structuring fats (often called“butters”) are generally solid at room temperature but melt sharplybetween 35-40° C.

High-SOS strains were obtained by three successive transformationsbeginning with strain S5100, a classically improved derivative, of awild type isolate of Prototheca moriformis, S376. Strain S5100 wastransformed with plasmid pSZ5654 to generate strain S8754, whichproduces an oil with increased stearic acid (C18:0) content, lowerpalmitic acid (C16:0) and reduced linoleic acid (C18:2cisΔ9,12) contentrelative to S5100. In turn, strain S8754 was transformed with plasmidpSZ5868 to generate strain S8813, which produces oil with higher C18:0,lower C16:0 and improved sn-2 selectivity compared to S8754. Finally,strain S8813 was transformed with plasmids pSZ6383 or pSZ6384 togenerate strains S9119, S9120 and S9121, producing oils rich in C18:0with reduced levels of C18:2cisΔ9,12 and improved sn-3 selectivity.

Construct Used for SAD2 Knockout in S5100

The first intermediate strains were prepared by transformation of strainS5100 with integrative plasmid pSZ5654(SAD2-1vD::PmKASII-1tp_PmKASII-1_FLAG-CvNR:CrTUB2-PmFAD2hpA-CvNR:PmHXT1-2v2-ScarMEL1-PmPGK::SAD2-1vE).The construct targeted ablation of allele 1 of the endogenousstearoyl-ACP desaturase 2 gene (SAD2), concomitant with expression ofthe PmKASII gene encoding P. moriformis β-keto-acyl-ACP synthase, and aRNAi hairpin sequence to down-regulate fatty acid desaturase (FAD2) geneexpression. Deletion of one allele of SAD2 reduced SAD activity,resulting in elevated levels of C18:0. Overexpression of PmKASIIstimulated elongation of C16:0 to C18:0, further increasing C18:0. FAD2is responsible for the conversion of C18:1cisΔ9 (oleic) to C18:2cisΔ9,12(linoleic) fatty acids, and RNAi of FAD2 resulted in decreased C18:2.Thus, the first intermediate strains had higher levels of C18:0 anddecreased C16:0 and C18:2 fatty acid levels relative to the S5100parent. The Saccharomyces carlsbergensis MEL1 gene, encoding a secretedmelibiase served as a selectable marker as part of plasmid pSZ5654,enabling the strain to grow on melibiose.

The sequence of the pSZ5654 transforming DNA is provided below. Relevantrestriction sites in the construct are indicated in lowercase, bold andunderlining and are 5′-3′ PmeI, SpeI, AscI, ClaI, SacI, AvrII, EcoRV,EcoRI, SpeI, BsiWI, XhoI, SacI, KpnI, SnaBI, BspQI and PmeI,respectively. PmeI sites delimit the 5′ and 3′ ends of the transformingDNA. Bold, lowercase sequences represent SAD2-1 5′ genomic DNA thatpermit targeted integration at the SAD2-1 locus via homologousrecombination. Proceeding in the 5′ to 3′ direction, bold, lowercasesequences represent SAD2-1 5′ genomic DNA sequences that permit targetedintegration at the FATA-1 locus via homologous recombination. Theinitiator ATG of the sequence encoding the P. moriformis KASII-1 transitpeptide (PmKASII-1tp) is indicated by uppercase, bold italics, and thePmKASII-1tp sequence located between the ATG and the AscI site isindicated with lowercase, underlined italics. The PmKASII-1 codingregion is indicated by lowercase italics. A sequence encoding a 3×FLAGtag fused to the C-terminus of PmKASII-1 is represented by uppercaseitalics, and the TGA terminator codon is indicated with uppercase, bolditalics. The Chlorella vulgaris nitrate reductase (NR) gene 3′ UTR isindicated by lowercase underlined text. A spacer sequence is representedby lowercase text. The C. reinhardtii TUB2 promoter, driving expressionof the PmFAD2hpA sequence is indicated by boxed text. Bold italicsdenote the PmFAD2hpA sequence followed by lowercase underlined textrepresenting C. vulgaris nitrate reductase 3′ UTR. A second spacersequence is represented by lowercase text. The P. moriformis HXT1promoter driving the expression of the S. carlbergensis MEL1 gene isindicated by boxed text. The initiator ATG and terminator TGA for MEL1gene are indicated by uppercase, bold italics while the coding region isindicated in lowercase italics. The P. moriformis PGK 3′ UTR isindicated by lowercase underlined text. The SAD2-1 3′ genomic regionindicated by bold, lowercase text.

Nucleotide sequence of transforming DNA contained in pSZ5654SEQ ID NO: 126  gtttaaacgccggtcaccacccgcatgctcgtactacagcgcacgcaccgcttcgtgatccaccgggtgaacgtagtcctcgacggaaacatctggttcgggcctcctgcttgcactcccgcccatgccgacaacctttctgctgttaccacgacccacaatgcaacgcgacacgaccgtgtgggactgatcggttcactgcacctgcatgcaattgtcacaagcgcttactccaattgtattcgtttgttttctgggagcagttgctcgaccgcccgcgtcccgcaggcagcgatgacgtgtgcgtggcctgggtgtttcgtcgaaaggccagcaaccctaaatcgcaggcgatccggagattgggatctgatccgagtttggaccagatccgccccgatgcggcacgggaactgcatcgactcggcgcggaacccagctttcgtaaatgccagattggtgtccgatacctggatttgccatcagcgaaacaagacttcagcagcgagcgtatttggcgggcgtgctaccagggttgcatacattgcccatttctgtctggaccgctttactggcgcagagggtgagttgatggggttggcaggcatcgaaacgcgcgtgcatggtgtgcgtgtctgttttcggctgcacgaattcaatagtcggatgggcgacggtagaattgggtgtggcgctcgcgtgcatgcctcgccccgtcgggtgtcatgaccgggactggaatcccccctcgcgaccatcttgctaacgctcccgactc

gccactacttcaacacccacctacccaccacctcccaccacaccatacaccacacctaatcccacatcacccacg ggcgcgcc gccgccgccgccgacgccaaccccgcccgccccgagcgccgcgtggtgatcaccggccagggcgtggtgacctccctgggccagaccatcgagcagttctactcctccctgctggagggcgtgtccggcatctcccagatccagaagttcgacaccaccggctacaccaccaccatcgccggcgagatcaagtccctgcagctggacccctacgtgcccaagcgctgggccaagcgcgtggacgacgtgatcaagtacgtgtacatcgccggcaagcaggccctggagtccgccggcctgcccatcgaggccgccggcctggccggcgccggcctggaccccgccctgtgcggcgtgctgatcggcaccgccatggccggcatgacctccttcgccgccggcgtggaggccctgacccgcggcggcgtgcgcaagatgaaccccttctgcatccccttctccatctccaacatgggcggcgccatgctggccatggacatcggcttcatgggccccaactactccatctccaccgcctgcgccaccggcaactactgcatcctgggcgccgccgaccacatccgccgcggcgacgccaacgtgatgctggccggcggcgccgacgccgccatcatcccctccggcatcggcggcttcatcgcctgcaaggccctgtccaagcgcaacgacgagcccgagcgcgcctcccgcccctgggacgccgaccgcgacggcttcgtgatgggcgagggcgccggcgtgctggtgctggaggagctggagcacgccaagcgccgcggcgccaccatcctggccgagctggtgggcggcgccgccacctccgacgcccaccacatgaccgagcccgacccccagggccgcggcgtgcgcctgtgcctggagcgcgccctggagcgcgcccgcctggcccccgagcgcgtgggctacgtgaacgcccacggcacctccacccccgccggcgacgtggccgagtaccgcgccatccgcgccgtgatcccccaggactccctgcgcatcaactccaccaagtccatgatcggccacctgctgggcggcgccggcgccgtggaggccgtggccgccatccaggccctgcgcaccggctggctgcaccccaacctgaacctggagaaccccgcccccggcgtggaccccgtggtgctggtgggcccccgcaaggagcgcgccgaggacctggacgtggtgctgtccaactccttcggcttcggcggccacaactcctgcgtgatcttccgcaagtacgacgagATGGACTACAAGGACCACGACGGCGACTACAA

cactctggacgctggtcgtgtgatggactgttgccgccacacttgctgccttgacctgtgaatatccctgccgcttttatcaaacagcctcagtgtgtttgatcttgtgtgtacgcgcttttgcgagttgctagctgcttgtgctatttgcgaataccacccccagcatccccttccctcgtttcatatcgcttgcatcccaaccgcaacttatctacgctgtcctgctatccctcagcgctgctcctgctcctgctcactgcccctcgcacagccttggtttgggctccgcctgtattctcctggtactgcaacctgtaaaccagcactgcaatgctgatgcacgggaagtagtgggatgggaacacaaatggagagctccgcgtctcgaacagagcgcgcagaggaacgctgaaggtctcgcctctgtcgcacctcagcgcggcatacaccacaataaccacctgacgaatgcgcttggttcttcgtccattagcgaagcgtccggttcacacacgtgccacgttggcg

cacactctggacgctggtcgtgtgatggactgttgccgccacacttgctgccttgacctgtgaatatccctgccgcttttatcaaacagcctcagtgtgtttgatcttgtgtgtacgcgcttttgcgagttgctagctgcttgtgctatttgcgaataccacccccagcatccccttccctcgtttcatatcgcttgcatcccaaccgcaacttatctacgctgtcctgctatccctcagcgctgctcctgctcctgctcactgcccctcgcacagccttggtttgggctccgcctgtattctcctggtactgcaacctgtaaaccagcactgcaatgctgatgcacgggaagtagtgggatgggaacacaaatggaaagctgtagagctcgatctaagtaagattcgaagcgctcgaccgtgccggacggactgcagccccatgtcgtagtgaccgccaatgtaagtgggctggcgtttccctgtacgtgagtcaacgtcactgcacgcgcaccaccctctcgaccggca

ctacaacggcctgggcctgacgccccagatgggctgggacaactggaacacgttcgcctgcgacgtctccgagcagctgctgctggacacggccgaccgcatctccgacctgggcctgaaggacatgggctacaagtacatcatcctggacgactgctggtcctccggccgcgactccgacggcttcctggtcgccgacgagcagaagttccccaacggcatgggccacgtcgccgaccacctgcacaacaactccttcctgttcggcatgtactcctccgcgggcgagtacacgtgcgccggctaccccggctccctgggccgcgaggaggaggacgcccagttcttcgcgaacaaccgcgtggactacctgaagtacgacaactgctacaacaagggccagttcggcacgcccgagatctcctaccaccgctacaaggccatgtccgacgccctgaacaagacgggccgccccatcttctactccctgtgcaactggggccaggacctgaccttctactggggctccggcatcgcgaactcctggcgcatgtccggcgacgtcacggcggagttcacgcgccccgactcccgctgcccctgcgacggcgacgagtacgactgcaagtacgccggcttccactgctccatcatgaacatcctgaacaaggccgcccccatgggccagaacgcgggcgtcggcggctggaacgacctggacaacctggaggtcggcgtcggcaacctgacggacgacgaggagaaggcgcacttctccatgtgggccatggtgaagtcccccctgatcatcggcgcgaacgtgaacaacctgaaggcctcctcctactccatctactcccaggcgtccgtcatcgccatcaaccaggactccaacggcatccccgccacgcgcgtctggcgctactacgtgtccgacacggacgagtacggccagggcgagatccagatgtggtccggccccctggacaacggcgaccaggtcgtggcgctgctgaacggcggctccgtgtcccgccccatgaacacgaccctggaggagatcttcttcgactccaacctgggctccaagaagctgacctccacctgggacatctacgacctgtgggcgaaccgcgtcgacaactccacggcgtccgccatcctgggccgcaacaagaccgccaccggcatcctgtacaacgccaccgagcagtcctacaaggacggcctgtccaagaacgacacccgcctgttcggccagaagatcggctccctgtcccccaacgcgatcctgaacacgaccgtccccgcccacggcatcgcgttct

ttactcttgaggaattgaacctttctcgcttgctggcatgtaaacattggcgcaattaattgtgtgatgaagaaagggtggcacaagatggatcgcgaatgtacgagatcgacaacgatggtgattgttatgaggggccaaacctggctcaatcttgtcgcatgtccggcgcaatgtgatccagcggcgtgactctcgcaacctggtagtgtgtgcgcaccgggtcgctttgattaaaactgatcgcattgccatcccgtcaactcacaagcctactctagctcccattgcgcactcgggcgcccggctcgatcaatgttctgagcggagggcgaagcgtcaggaaatcgtctcggcagctggaagcgcatggaatgcggagcggagatcgaatcaggatccttagggagcgacgagtgtgcgtgcggggctggcgggagtgggacgccctcctcgctcctctctgttctgaacggaacaatcggccaccccgcgctacgcgccacgcatcgagcaacgaagaaaaccccccgatgataggttgcggtggctgccgggatatagatccggccgcacatcaaagggcccctccgccagagaagaagctcctttcccagcagactccttctgctgccaaaacacttctctgtccacagcaacaccaaaggatgaacagatcaacttgcgtctccgcgtagcttcctcggctagcgtgcttgcaacaggtccctgcactattatcttcctgctttcctctgaattatgcggcaggcgagcgctcgctctggcgagcgctccttcgcgccgccctcgctgatcgagtgtacagtcaatgaatggtcctgggcgaagaacgagggaatttgtgggtaaaacaagcatcgtctctcaggccccggcgcagtggccgttaaagtccaagaccgtgaccaggcagcgcagcgcgtccgtgtgcgggccctgcctggcggctcggcgtgccaggctcgagagcagctccctcaggtcgccttggacggcctctgcgaggccggtgagggcctgcaggagcgcctcgagcgtggcagtggcggtcgtatccgggtcgccggtcaccgcctgcgactcgccatccgaagagcg tttaaac

Construct pSZ5654 was transformed into S5100. Primary transformants wereclonally purified and screened under standard lipid productionconditions at pH 5. Integration of pSZ5654 at the SAD2-1 locus wasverified by DNA blot analysis. The fatty acid profiles and lipid titersof lead strains were assayed in 50-mL shake flasks (Table 21). S8754 wasselected as the lead strain for additional rounds of geneticengineering. As shown in Table 21, C16:0 decreased from 17.6% to lessthan 6%, C18:0 increased from 4.3% to about 28%, C18:2 decreased from5.8% to 1.3%.

TABLE 21 Fatty acid profiles of SAD2-1 ablation strains. Sample ID S5100S8741 S8742 S8743 S8744 S8745 S8746 S8752 S8753 S8754 C14:0 0.7 0.6 0.60.6 0.6 0.6 0.6 0.6 0.6 0.6 C16:0 17.6 5.9 5.9 5.8 5.9 5.9 5.9 5.9 5.85.9 C16:1 cis-9 0.4 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.1 C18:0 4.3 28.228.1 27.7 27.8 27.4 28.2 28.3 28.3 28.1 C18:1 69.8 60.1 60.2 60.6 60.560.9 60.0 60.0 60.0 60.0 C18:2 5.8 1.3 1.3 1.3 1.3 1.3 1.3 1.3 1.2 1.3C18:3 α 0.5 0.3 0.3 0.3 0.3 0.3 0.3 0.3 0.3 0.3 C20:0 0.3 2.2 2.2 2.22.2 2.2 2.2 2.2 2.2 2.2 saturates 23.2 37.5 37.5 37.1 37.2 36.8 37.737.7 37.7 37.6 lipid (g/L) 13.5 12.8 12.5 12.5 12.5 12.3 12.3 12.3 12.412.3

Construct Used for FATA-1 Knockout in S8754

The second intermediate strains were prepared by transformation ofstrain S8754 with integrative plasmid pSZ5868(FATA-1vB::CpSAD1tp_GarmFATA1(G108A)_FLAG-PmSAD2-1:PmG3PDH-1-TcLPAT2-PmATP:CrTUB2-ScSUC2-PmPGH::FATA-1vC).This construct targeted ablation of allele 1 of the endogenous fattyacyl-ACP thioesterase gene (FATA-1), and contained expression modulesfor GarmFATA1(G108A), encoding a variant of the Garcinia mangostanaFATA1 thioesterase with improved activity, and TcLPAT2 encoding theTheobroma cacao lysophosphatidic acid acyltransferase (LPAAT). Deletionof one copy of FATA-1 reduced endogenous thioesterase activity, furtherreducing C16:0 accumulation. Expression of GarmFATA1(G108A) stimulatedC18:0-ACP hydrolysis, further increasing C18:0. TcLPAT2 had superiorspecificity for transfer of C18:1 to the sn-2 position oftriacylglycerides than the endogeneous LPAAT, leading to reducedaccumulation of trisaturates. The second intermediate strains hadincreased C18:0 and lower C16:0 compared their parent, S8754. The S.cerevisiae SUC2 gene encoding a secreted sucrose invertase, served as aselectable marker as part of plasmid pSZ5868 and enabled the strain togrow on sucrose.

The sequence of the pSZ5868 transforming DNA is provided below. Relevantrestriction sites in the construct are indicated in lowercase, bold andunderlining and are 5′-3′ BspQI, PmeI, SpeI, AscI, ClaI, SacI, AvrII,NdeI, NsiI, AfIII, KpnI, XbaI, MfeI, BamHI, BspQI and PmeI,respectively. BspQI and PmeI sites delimit the 5′ and 3′ ends of thetransforming DNA. Proceeding in the 5′ to 3′ direction, bold, lowercasesequences represent FATA-1 5′ genomic DNA that permit targetedintegration at the FATA-1 locus via homologous recombination. Theinitiator ATG of the sequence encoding the C. protothecoides SAD1transit peptide (CpSAD1tp) is indicated by uppercase, bold italics, andthe remainder of the CpSAD1tp sequence located between the ATG and theAscI site is indicated with lowercase, underlined italics. TheGarmFATA1(G108A) coding region is indicated by lowercase italics. Asequence encoding a 3×FLAG tag fused to the C-terminus ofGarmFATA1(G108A) is represented by uppercase italics, and the TGAterminator codon is indicated with uppercase, bold italics. The P.moriformis SAD2-1 3′ UTR is indicated by lowercase underlined text. Aspacer sequence is represented by lowercase text. The P. moriformisG3PDH-1 promoter, driving expression of the TcLPAT2 sequence isindicated by boxed text. The initiator ATG and terminator TGA codons ofthe TcLPAT2 gene are indicated by uppercase, bold italics, while theremainder of the coding region is represented with italics. Lowercaseunderlined text represents the P. moriformis ATP 3′ UTR. A second spacersequence is represented by lowercase text. The C. reinhardtii TUB2promoter driving the expression of the S. cerevisiae SUC2 gene isindicated by boxed text. The initiator ATG and terminator TGA for SUC2are indicated by uppercase, bold italics while the coding region isindicated in lowercase italics. The P. moriformis PGH 3′ UTR isindicated by lowercase underlined text. The FATA-1 3′ genomic regionindicated by bold, lowercase text.

Nucleotide sequence of transforming DNA contained in pSZ5868SEQ ID NO: 127  gaagagc gcccaat gtttaaacctcttttgctgcgtctcctcaggcttgggggcctccttgggcttgggtgccgccatgatctgcgcgcatcagagaaacgttgctggtaaaaaggagcgcccggctgcgcaatatatatataggcatgccaacacagcccaacctcactcgggagcccgtcccaccacccccaagtcgcgtgccttgacggcatactgctgcagaagcttcatgagaatgatgccgaacaagaggggcacgaggacccaatcccggacatccttgtcgataatgatctcgtgagtccccatcgtccgcccgacgctccggggagcccgccgatgctcaagacgagagggccctcgaccaggaggggctggcccgggcgggcactggcgtcgaaggtgcgcccgtcgttcgcctgcagtcctatgccacaaaacaagtcttctgacggggtgcgtttgctcccgtgcgggcaggcaacagaggtattcaccctggtcatggggagatcggcgatcgagctgggataagagatacggtcccgcgcaaggatcgctcatcctggtctgagccggacagtcattctggcaagcaatgacaacttgtcaggaccggaccgtgccatatatttctcacctagcgccgcaaaacctaacaatttgggagtcactgtgccactgagttcgactggtagctgaatggagtcgctgctccactaaacgaattgtcagcaccgccagccggccgaggacccgagtcata

ggcgggctccgggccccggcgcccagcgaggcccctccccgtgcgcg ggcgcgccatccccccccgcatcatcgtggtgtcctcctcctcctccaaggtgaaccccctgaagaccgaggccgtggtgtcctccggcctggccgaccgcctgcgcctgggctccctgaccgaggacggcctgtcctacaaggagaagttcatcgtgcgctgctacgaggtgggcatcaacaagaccgccaccgtggagaccatcgccaacctgctgcaggaggtgggctgcaaccacgcccagtccgtgggctactccaccgccggcttctccaccacccccaccatgcgcaagctgcgcctgatctgggtgaccgcccgcatgcacatcgagatctacaagtaccccgcctggtccgacgtggtggagatcgagtcctggggccagggcgagggcaagatcggcacccgccgcgactggatcctgcgcgactacgccaccggccaggtgatcggccgcgccacctccaagtgggtgatgatgaaccaggacacccgccgcctgcagaaggtggacgtggacgtgcgcgacgagtacctggtgcactgcccccgcgagctgcgcctggccttccccgaggagaacaactcctccctgaagaagatctccaagctggaggacccctcccagtactccaagctgggcctggtgccccgccgcgccgacctggacatgaaccagcacgtgaacaacgtgacctacatcggctgggtgctggagtccatgccccaggagatcatcgacacccacgagctgcagaccatcaccctggactaccgccgcgagtgccagcacgacgacgtggtggactccctgacctcccccgagccctccgaggacgccgaggccgtgttcaaccacaacggcaccaacggctccgccaacgtgtccgccaacgaccacggctgccgcaacttcctgcacctgctgcgcctgtccggcaacggcctggagatcaaccgcggccgcaccgagtggcgcaagaagcccacccgcATGGACTACAAGGACCACGACGGCG

gcggggctggcgggagtgggacgccctcctcgctcctctctgttctgaacggaacaatcggccaccccgcgctacgcgccacgcatcgagcaacgaagaaaaccccccgatgataggttgcggtggctgccgggatatagatccggccgcacatcaaagggcccctccgccagagaagaagctcctttcccagcagactccttctgctgccaaaacacttctctgtccacagcaacaccaaaggatgaacagatcaacttgcgtctccgcgtagcttcctcggctagcgtgcttgcaacaggtccctgcactattatcttcctgctttcctctgaattatgcggcaggcgagcgctcgctctggcgagcgctccttcgcgccgccctcgctgatcgagtgtacagtcaatgaatggtgagctccgcgtctcgaacagagcgcgcagaggaacgctgaaggtctcgcctctgtcgcacctcagcgcggcatacaccacaataaccacctgacgaatgcgcttggttcttcgtccattagcgaagcgtccggttcacacacgtgccacgttggcgaggtggcaggtgacaatgatcggtggagctgatggt

tcgccgccgccgccgtgatcgtgcccctgggcctgctgttcttcatctccggcctggtggtgaacctgatccaggccctgtgcttcgtgctgatccgccccctgtccaagaacacctaccgcaagatcaaccgcgtggtggccgagctgctgtggctggagctgatctggctggtggactggtgggccggcgtgaagatcaaggtgttcatggaccccgagtccttcaacctgatgggcaaggagcacgccctggtggtggccaaccaccgctccgacatcgactggctggtgggctggctgctggcccagcgctccggctgcctgggctccgccctggccgtgatgaagaagtcctccaagttcctgcccgtgatcggctggtccatgtggttctccgagtacctgttcctggagcgctcctgggccaaggacgagaacaccctgaaggccggcctgcagcgcctgaaggacttcccccgccccttctggctggccttcttcgtggagggcacccgcttcacccaggccaagttcctggccgcccaggagtacgccgcctcccagggcctgcccatcccccgcaacgtgctgatcccccgcaccaagggcttcgtgtccgccgtgtcccacatgcgctccttcgtgcccgccatctacgacatgaccgtggccatccccaagtcctccccctcccccaccatgctgcgcctgttcaagggccagccctccgtggtgcacgtgcacatcaagcgctgcctgatgaaggagctgcccgagaccgacgaggccgtggcccagtggtgcaaggacatgttcgtggagaaggacaagctgctggacaagcacatcgccgaggacaccactccgaccagcccatgcaggacctgggccgccccatcaagtccctgctggtggtggcctcctgggcctgcctgatggcctacggcgccctgaagttcctgcagtgctcctccctgctgtcctcctggaagggcatcgccacttcctggtgggcctggccatcgtgaccatcctgatgcacatcctgatcctgttctcccagtccgagcgctccacccccgccaaggtgg

agggtggtcgactcgttggaggtgggtgtttttttttatcgagtgcgcggcgcggcaaacgggtccctttttatcgaggtgttcccaacgccgcaccgccctcttaaaacaacccccaccaccacttgtcgaccttctcgtttgttatccgccacggcgccccggaggggcgtcgtctggccgcgcgggcagctgtatcgccgcgctcgctccaatggtgtgtaatcttggaaagataataatcgatggatgaggaggagagcgtgggagatcagagcaaggaatatacagttggcacgaagcagcagcgtactaagctgtagcgtgttaagaaagaaaaactcgctgttaggctgtattaatcaaggagcgtatcaataattaccgaccctatacctttatctccaacccaatcgcggcttaaggatctaagtaagattcgaagcgctcgaccgtgccggacggactgcagccccatgtcgtagtgaccgccaatgtaagtgggctggcgtttccctgtacgtgagtcaacgtcactgcacgcgcaccaccctctcgaccggcaggaccaggcatcgcgagatacagcgcgagccagacacggagtg

ccgaccgccccctggtgcacttcacccccaacaagggctggatgaacgaccccaacggcctgtggtacgacgagaaggacgccaagtggcacctgtacttccagtacaacccgaacgacaccgtctgggggacgcccttgactggggccacgccacgtccgacgacctgaccaactgggaggaccagcccatcgccatcgccccgaagcgcaacgactccggcgccttctccggctccatggtggtggactacaacaacacctccggcttcttcaacgacaccatcgacccgcgccagcgctgcgtggccatctggacctacaacaccccggagtccgaggagcagtacatctcctacagcctggacggcggctacaccttcaccgagtaccagaagaaccccgtgctggccgccaactccacccagaccgcgacccgaaggtcttctggtacgagccctcccagaagtggatcatgaccgcggccaagtcccaggactacaagatcgagatctactcctccgacgacctgaagtcctggaagctggagtccgcgttcgccaacgagggcacctcggctaccagtacgagtgccccggcctgatcgaggtccccaccgagcaggaccccagcaagtcctactgggtgatgttcatctccatcaaccccggcgccccggccggcggctccttcaaccagtacttcgtcggcagcttcaacggcacccacttcgaggccttcgacaaccagtcccgcgtggtggacttcggcaaggactactacgccctgcagaccttcttcaacaccgacccgacctacgggagcgccctgggcatcgcgtgggcctccaactgggagtactccgccacgtgcccaccaacccctggcgctcctccatgtccctcgtgcgcaagttctccctcaacaccgagtaccaggccaacccggagacggagctgatcaacctgaaggccgagccgatcctgaacatcagcaacgccggcccctggagccggttcgccaccaacaccacgttgacgaaggccaacagctacaacgtcgacctgtccaacagcaccggcaccctggagttcgagctggtgtacgccgtcaacaccacccagacgatctccaagtccgtgttcgcggacctctccctctggttcaagggcctggaggaccccgaggagtacctccgcatgggcttcgaggtgtccgcgtcctccttcttcctggaccgcgggaacagcaaggtgaagttcgtgaaggagaacccctacttcaccaaccgcatgagcgtgaacaaccagcccttcaagagcgagaacgacctgtcctactacaaggtgtacggcttgctggaccagaacatcctggagctgtacttcaacgacggcgacgtcgtgtccaccaacacctacttcatgaccaccgggaacgccctgggctccgtgaacatgacgacgggggtggacaacctgttctacatcgaca

cgaaacaagcccctggagcatgcgtgcatgatcgtctctggcgccccgccgcgcggtttgtcgccctcgcgggcgccgcggccgcgggggcgcattgaaattgttgcaaaccccacctgacagattgagggcccaggcaggaaggcgttgagatggaggtacaggagtcaagtaactgaaagtttttatgataactaacaacaaagggtcgtttctggccagcgaatgacaagaacaagattccacatttccgtgtagaggcttgccatcgaatgtgagcgggcgggccgcggacccgacaaaacccttacgacgtggtaagaaaaacgtggcgggcactgtccctgtagcctgaagaccagcaggagacgatcggaagcatcacagcacaggatcctgaggacagggtggttggctggatggggaaacgctggtcgcgggattcgatcctgctgcttatatcctccctggaagcacacccacgactctgaagaagaaaacgtgcacacacacaacccaaccggccgaatatttgcttccttatcccgggtccaagagagactgcgatgcccccctcaatcagcatcctcctccctgccgcttcaatcttccctgcttgcctgcgcccgcggtgcgccgtctgcccgcccagtcagtcactcctgcacaggccccttgtgcgcagtgctcctgtaccctttaccgctccttccattctgcgaggccccctattgaatgtattcgttgcctgtgtggccaagcgggctgctgggcgcgccgccgtcgggcagtgctcggcgactttggcggaagccgattgttcttctgtaagccacgcgcttgctgctttgggaagagaagggggggggtactgaatggatgaggaggagaaggaggggtattggtattatctgagttggggaggcagggagagttggaaaatgtaagtggcacgacgggcaaggagaatggtgagcatgtgcatggtgatgtcgttggtcgaggacgatcctgcacgcgtgtatctgatgtagaatacggcaatcaccctagtctacatctataccttctccgtataacgccctttccaaatgccctcccgtttctctcctattcttgatccacatgatgaccctggcactatttcaagggctggag aagagcgtttaaac

Construct pSZ5868 was transformed into S8754. Primary transformants wereclonally purified and screened under standard lipid productionconditions at pH 5. Integration of pSZ5868 at the FATA-1 locus wasverified by DNA blot analysis. The fatty acid profiles and lipid titersof lead strains were assayed in 50-mL shake flasks (Table 22). S8813 wasselected as the lead strain for the final round of genetic engineering.As shown in Table 22 as compared to strain S8754, C16:0 decreased from5.9% to 3.4%, and C18:0 increased from 27.3% to about 45%. C18:2increased slightly from 1.3% to about 1.6% due to the activity of the T.cacao LPAAT.

TABLE 22 Fatty acid profiles of FATA-1 ablation strains. Strain 5510058754 58813 58814 C14:0 0.7 0.6 0.5 0.5 C16:0 18.8 5.9 3.4 3.4 C16:1cis-9 0.5 0.0 0.0 0.0 C18:0 4.0 27.3 45.3 44.8 C18:1 68.3 60.9 45.9 46.3C18:2 6.3 1.3 1.5 1.6 C18:3 α 0.6 0.3 0.3 0.3 C20:0 0.3 2.4 2.0 2.1saturates 24.2 37.0 52.0 51.5 lipid (g/L) 12.7 11.9 11.9 11.9

Constructs Used for FAD2 Knockout in S8813

The high-SOS strains were generated by transformation of strain S8813with integrative plasmid pSZ6383(FAD2-1vA::PmLDH1-AtTHIC-PmHSP90:PmSAD2-2v2-TcDGAT1-CvNR:PmSAD2-1v3-CpSAD1tp_GarmFATA1(G108A)_FLAG-PmSAD2-1::FAD2-1vB),plasmid pSZ6384(FAD2-1vA::PmLDH1-AtTHIC-PmHSP90:PmSAD2-2v2-TcDGAT2-CvNR:PmSAD2-1v3-CpSAD1tp_GarmFATA1(G108A)_FLAG-PmSAD2-1::FAD2-1vB),or plasmid pSZ6377 (FAD2-1vA::PmLDH1-AtTHIC-PmHSP90:PmSAD2-1v3-CpSAD1tp_GarmFATA1(G108A)_FLAG-PmSAD2-1::FAD2-1vB). Theseconstructs targeted ablation of allele 1 of the endogenous fatty aciddesaturase 2 gene (FAD2-1), and contained expression modules for asecond copy of GarmFATA1(G108A), and either TcDGAT1 encoding theTheobroma cacao diacylglycerol O-acyltransferase 1 (pSZ6383) or TcDGAT2encoding the Theobroma cacao diacylglycerol O-acyltransferase 2(pSZ6384). Deletion of one allele of FAD2 further reduced C18:2accumulation. Expression of GarmFATA1(G108A) stimulated C18:0-ACPhydrolysis, further increasing C18:0. TcDGAT1 and TcDGAT2 had superiorspecificity for transfer of C18:0 to the sn-3 position oftriacylglycerides than the endogeneous DGAT, leading to an increase inC18:0 and lipid titer, and a reduction in trisaturated TAGs. The finalstrains had higher C18:0, lower C16:0 and lower C18:2 than their parent,S8813. The Arabidopsis thaliana THIC gene (AtTHIC) catalyzes theconversion of 5-aminoimidazole ribotide (AIR) to4-amino-5-hydroxymethylpyrimidine (HMP), providing the pyrimidine ringstructure for the biosynthesis of thiamine. AtTHIC served as aselectable marker as part of plasmids pSZ6383 and pSZ6384, allowing thestrains to grow in the absence of exogenous thiamine.

The sequence of the pSZ6383 transforming DNA is provided below. Relevantrestriction sites in the construct are indicated in lowercase, bold andunderlined text, and are 5′-3′ BspQI, KpnI, XbaI, SnaBI, BamHI, AvrII,SpeI, ClaI, AflII, EcoRI, SpeI, AscI, ClaI, SacI and BspQ I,respectively. BspQI sites delimit the 5′ and 3′ ends of the transformingDNA. Proceeding in the 5′ to 3′ direction, bold, lowercase sequencesrepresent FAD2-1 5′ genomic DNA that permits targeted integration at theFAD2-1 locus via homologous recombination. The P. moriformis LDH1promoter driving the expression of the Arabidopsis thaliana THIC gene isindicated by boxed text. The initiator ATG and terminator TGA for AtTHICare indicated by uppercase, bold italics while the coding region isindicated in lowercase italics. The P. moriformis HSP90 3′ UTR isindicated by lowercase underlined text. A spacer sequence is representedby lowercase text. The P. moriformis SAD2-2 promoter, driving expressionof the TcDGAT1 sequence is indicated by boxed text. The initiator ATGand terminator TGA codons of the TcDGAT1 gene are indicated byuppercase, bold italics, while the remainder of the coding region isrepresented with italics. Lowercase underlined text represents the C.vulgaris NR 3′ UTR. A second spacer sequence is represented by lowercasetext. The P. moriformis SAD2-1 promoter, indicated by boxed italicizedtext, is utilized to drive the expression of the G. mangostana FATA1gene. The initiator ATG of the sequence encoding the C. protothecoidesSAD1 transit peptide (CpSAD1tp) is indicated by uppercase, bold italics,and the remainder of the CpSAD1tp sequence located between the ATG andthe AscI site is indicated with lowercase, underlined italics. TheGarmFATA1(G108A) coding region is indicated by lowercase italics. Asequence encoding a 3×FLAG tag fused to the C-terminus ofGarmFATA1(G108A) is represented by uppercase italics, and the TGAterminator codon is indicated with uppercase, bold italics. The P.moriformis SAD2-1 3′ UTR is indicated by lowercase underlined text. TheFAD2-1 3′ genomic region is indicated by bold, lowercase text.

Nucleotide sequence of transforming DNA contained in pSZ6383SEQ ID NO: 128  gctcttcgcgaaggtcattttccagaacaacgaccatggcttgtcttagcgatcgctcgaatgactgctagtgagtcgtacgctcgacccagtcgctcgcaggagaacgcggcaactgccgagcttcggcttgccagtcgtgactcgtatgtgatcaggaatcattggcattggtagcattataattcggcttccgcgctgtttatgggcatggcaatgtctcatgcagtcgaccttagtcaaccaattctgggtggccagctccgggcgaccgggctccgtgtcgccgggcaccacctcctgccatgagtaacagggccgccctctcctcccgacgttggccaactgaataccgtgtcttggggccctacatgatgggctgcctagtcgggcgggacgcgcaactgcccgcgcaatctgggacgtggtctgaatcctccaggcgggtttccccgagaaagaaagggtgccgatttcaaagcagagccatgtgccgggccctgtggcctgtgttggcgcctatgtagtcaccccccctcacccaattgtcgccagtttgcgcaatccataaactcaaaactgcagcttctgagctgcgctgttcaa

ctgatgtccgtggtctgcaacaacaagaaccactccgcccgccccaagctgcccaactcctccctgctgcccggcttcgacgtggtggtccaggccgcggccacccgcttcaagaaggagacgacgaccacccgcgccacgctgacgttcgacccccccacgaccaactccgagcgcgccaagcagcgcaagcacaccatcgacccctcctcccccgacttccagcccatcccctccttcgaggagtgcttccccaagtccacgaaggagcacaaggaggtggtgcacgaggagtccggccacgtcctgaaggtgcccttccgccgcgtgcacctgtccggcggcgagcccgccttcgacaactacgacacgtccggcccccagaacgtcaacgcccacatcggcctggcgaagctgcgcaaggagtggatcgaccgccgcgagaagctgggcacgccccgctacacgcagatgtactacgcgaagcagggcatcatcacggaggagatgctgtactgcgcgacgcgcgagaagctggaccccgagttcgtccgctccgaggtcgcgcggggccgcgccatcatcccctccaacaagaagcacctggagctggagcccatgatcgtgggccgcaagttcctggtgaaggtgaacgcgaacatcggcaactccgccgtggcctcctccatcgaggaggaggtctacaaggtgcagtgggccaccatgtggggcgccgacaccatcatggacctgtccacgggccgccacatccacgagacgcgcgagtggatcctgcgcaactccgcggtccccgtgggcaccgtccccatctaccaggcgctggagaaggtggacggcatcgcggagaacctgaactgggaggtgttccgcgagacgctgatcgagcaggccgagcagggcgtggactacttcacgatccacgcgggcgtgctgctgcgctacatccccctgaccgccaagcgcctgacgggcatcgtgtcccgcggcggctccatccacgcgaagtggtgcctggcctaccacaaggagaacttcgcctacgagcactgggacgacatcctggacatctgcaaccagtacgacgtcgccctgtccatcggcgacggcctgcgccccggctccatctacgacgccaacgacacggcccagttcgccgagctgctgacccagggcgagctgacgcgccgcgcgtgggagaaggacgtgcaggtgatgaacgagggccccggccacgtgcccatgcacaagatccccgagaacatgcagaagcagctggagtggtgcaacgaggcgcccttctacaccctgggccccctgacgaccgacatcgcgcccggctacgaccacatcacctccgccatcggcgcggccaacatcggcgccctgggcaccgccctgctgtgctacgtgacgcccaaggagcacctgggcctgcccaaccgcgacgacgtgaaggcgggcgtcatcgcctacaagatcgccgcccacgcggccgacctggccaagcagcacccccacgcccaggcgtgggacgacgcgctgtccaaggcgcgcttcgagttccgctggatggaccagttcgcgctgtccctggaccccatgacggcgatgtccttccacgacgagacgctgcccgcggacggcgcgaaggtcgcccacttctgctccatgtgcggccccaagttctgctccatgaagatcacggaggacatccgcaagtacgccgaggagaacggctacggctccgccgaggaggccatccgccagggcatggacgccatgtccgaggagttcaacatcgccaagaagacgatctccggcgagcagcacggcgaggtcggcggcgagatctacctgcccgagtcctacgtc

cgcacgcatccaacgaccgtatacgcatcgtccaatgaccgtcggtgtcctctctgcctccgttttgtgagatgtctcaggcttggtgcatcctcgggtggccagccacgttgcgcgtcgtgctgcttgcctctcttgcgcctctgtggtactggaaaatatcatcgaggcccgtttttttgctcccatttcctttccgctacatcttgaaagcaaacgacaaacgaagcagcaagcaaagagcacgaggacggtgaacaagtctgtcacctgtatacatctatttccccgcgggtgcacctactctctctcctgccccggcagagtcagctgccttacgtgacggatcccgcgtctcgaacagagcgcgcagaggaacgctgaaggtctcgcctctgtcgcacctcagcgcggcatacaccacaataaccacctgacgaatgcgcttggttcttcgtccattagcgaagcgtccggttcacacacgtgccacgttggcgaggtggcaggtgacaatgatcggtgga

cgagatcctgggctccaccgccaccgtgacctcctcctcccactccgactccgacctgaacctgctgtccatccgccgccgcacctccaccaccgccgccgcccgcgcccccgaccgcgacgactccggcaacggcgaggccgtggacgaccgcgaccgcgtggagtccgccaacctgatgtccaacgtggccgagaacgccaacgagatgcccaactcctccgacacccgcttcacctaccgcccccgcgtgcccgcccaccgccgcatcaaggagtcccccctgtcctccggcgccatcttcaagcagtcccacgccggcctgttcaacctgtgcatcgtggtgctggtggccgtgaactcccgcctgatcatcgagaacctgatgaagtacggctggctgatccgctccggcttctggttctcctcccgctccctgtccgactggcccctgttcatgtgctgcctgaccctgcccatcttccccctggccgccttcgtggtggagaagctggtgcagcgcaactacatctccgagcccgtggtggtgttcctgcacgccatcatctccaccaccgccgtgctgtaccccgtgatcgtgaacctgcgctgcgactccgccttcctgtccggcgtggccctgatgctgttcgcctgcatcgtgtggctgaagctggtgtcctacgcccacaccaacaacgacatgcgcgccctggccaagtccgccgagaagggcgacgtggacccctcctacgacgtgtccttcaagtccctggcctacttcatggtggcccccaccctgtgctaccagcagtcctacccccgcacccccgccgtgcgcaagtcctgggtggtgcgccagttcatcaagctgatcgtgttcaccggcctgatgggcttcatcatcgagcagtacatcaaccccatcgtgcagaactcccagcaccccctgaagggcaacctgctgtacgccatcgagcgcgtgctgaagctgtccgtgcccaacctgtacgtgtggctgtgcatgttctactgcttcttccacctgtggctgaacatcctggccgagctgctgcgcttcggcgaccgcgagttctacaaggactggtggaacgccaagaccgtggaggagtactggcgcatgtggaacatgcccgtgcacaagtggatggtgcgccacatctacttcccctgcctgcgcaacggcatccccaagggcgtggccatcgtgatcgccttcctggtgtccgccgtgttccacgagctgtgcatcgccgtgccctgccacatgttcaagctgtgggccttcatcggcatcatgttccaggtgcccctggtgctgatcaccaactacctgcaggacaagttccgctcctccatggtgggcaacatgatcttctggttcatcttctccatcctgggccagcccatgtgcgtgctgctgt

gacacactctggacgctggtcgtgtgatggactgttgccgccacacttgctgccttgacctgtgaatatccctgccgcttttatcaaacagcctcagtgtgtttgatcttgtgtgtacgcgcttttgcgagttgctagctgcttgtgctatttgcgaataccacccccagcatccccttccctcgtttcatatcgcttgcatcccaaccgcaacttatctacgctgtcctgctatccctcagcgctgctcctgctcctgctcactgcccctcgcacagccttggtttgggctccgcctgtattctcctggtactgcaacctgtaaaccagcactgcaatgctgatgcacgggaagtagtgggatgggaacacaaatggacttaaggatctaagtaagattcgaagcgctcgaccgtgccggacggactgcagccccatgtcgtagtgaccgccaatgtaagtgggctggcgtttccctgtacgtgagtcaacgtcactgcacgcgcaccaccctctcgaccggcaggaccaggcatcgcgagatacagcgcgagccagacacggagtgccgagctatgcgcacgctccaactagatatcatgtggatgatgagcat

aatgcccactgcggcgacctgcgtcgctcggcgggctccgggccccggcgcccagcgaggcccctccccgtgcgcgggcgcgc catccccccccgcatcatcgtggtgtcctcctcctcctccaaggtgaaccccctgaagaccgaggccgtggtgtcctccggcctggccgaccgcctgcgcctgggctccctgaccgaggacggcctgtcctacaaggagaagttcatcgtgcgctgctacgaggtgggcatcaacaagaccgccaccgtggagaccatcgccaacctgctgcaggaggtgggctgcaaccacgcccagtccgtgggctactccaccgccggcttctccaccacccccaccatgcgcaagctgcgcctgatctgggtgaccgcccgcatgcacatcgagatctacaagtaccccgcctggtccgacgtggtggagatcgagtcctggggccagggcgagggcaagatcggcacccgccgcgactggatcctgcgcgactacgccaccggccaggtgatcggccgcgccacctccaagtgggtgatgatgaaccaggacacccgccgcctgcagaaggtggacgtggacgtgcgcgacgagtacctggtgcactgcccccgcgagctgcgcctggccttccccgaggagaacaactcctccctgaagaagatctccaagctggaggacccctcccagtactccaagctgggcctggtgccccgccgcgccgacctggacatgaaccagcacgtgaacaacgtgacctacatcggctgggtgctggagtccatgccccaggagatcatcgacacccacgagctgcagaccatcaccctggactaccgccgcgagtgccagcacgacgacgtggtggactccctgacctcccccgagccctccgaggacgccgaggccgtgttcaaccacaacggcaccaacggctccgccaacgtgtccgccaacgaccacggctgccgcaacttcctgcacctgctgcgcctgtccggcaacggcctggagatcaaccgcggccgcaccgagtggcgcaagaagcccacccgcATGGACTACAAGGACCACGACGGCGACTACAAGGACCACGACATCGACTACAAGGACGACGACGACAA

tcggccaccccgcgctacgcgccacgcatcgagcaacgaagaaaaccccccgatgataggttgcggtggctgccgggatatagatccggccgcacatcaaagggcccctccgccagagaagaagctcctttcccagcagactccttctgctgccaaaacacttctctgtccacagcaacaccaaaggatgaacagatcaacttgcgtctccgcgtagcttcctcggctagcgtgcttgcaacaggtccctgcactattatcttcctgctttcctctgaattatgcggcaggcgagcgctcgctctggcgagcgctccttcgcgccgccctcgctgatcgagtgtacagtcaatgaatggtgagctcctcactcagcgcgcctgcgcggggatgcggaacgccgccgccgccttgtcttttgcacgcgcgactccgtcgcttcgcgggtggcacccccattgaaaaaaacctcaattctgtttgtggaagacacggtgtacccccaaccacccacctgcacctctattattggtattattgacgcgggagcgggcgttgtactctacaacgtagcgtctctggttttcagctggctcccaccattgtaaattcttgctaaaatagtgcgtggttatgtgagaggtatggtgtaacagggcgtcagtcatgttggttttcgtgctgatctcgggcacaaggcgtcgtcgacgtgacgtgcccgtgatgagagcaataccgcgctcaaagccgacgcatggcctttactccgcactccaaacgactgtcgctcgtatttttcggatatctattttttaagagcgagcacagcgccgggcatgggcctgaaaggcctcgcggccgtgctcgtggtgggggccgcgagcgcgtggggcatcgcggcagtgcaccaggcgcagacggaggaacgcatggtgagtgcgcatcacaagatgcatgtcttgttgtctgtactataatgctagagcatcaccaggggcttagtcatcgcacctgctttggtcattacagaaattgcacaagggcgtcctccgggatgaggagatgtaccagctcaagctggagcggcttcgagccaagcaggagcgcggcgcatgacgacctacccacatgc gaagagc

The sequence of the pSZ6384 transforming DNA is provided below. Relevantrestriction sites in the construct are indicated in lowercase, bold andunderlined text, and are 5′-3′ BspQI, KpnI, XbaI, SnaBI, BamHI, AvrII,SpeI, ClaI, AfIII, EcoRI, SpeI, AscI, ClaI, SacI and BspQ I,respectively. BspQI sites delimit the 5′ and 3′ ends of the transformingDNA. Proceeding in the 5′ to 3′ direction, bold, lowercase sequencesrepresent FAD2-1 5′ genomic DNA that permits targeted integration at theFAD2-1 locus via homologous recombination. The P. moriformis LDH1promoter driving the expression of the Arabidopsis thaliana THIC gene isindicated by boxed text. The initiator ATG and terminator TGA for AtTHICare indicated by uppercase, bold italics while the coding region isindicated in lowercase italics. The P. moriformis HSP90 3′ UTR isindicated by lowercase underlined text. A spacer sequence is representedby lowercase text. The P. moriformis SAD2-2 promoter, driving expressionof the TcDGAT2 sequence is indicated by boxed text. The initiator ATGand terminator TGA codons of the TcDGAT2 gene are indicated byuppercase, bold italics, while the remainder of the coding region isrepresented with italics. Lowercase underlined text represents the C.vulgaris NR 3′ UTR. A second spacer sequence is represented by lowercasetext. The P. moriformis SAD2-1 promoter, indicated by boxed italicizedtext, is utilized to drive the expression of the G. mangostana FATA1gene. The initiator ATG of the sequence encoding the C. protothecoidesSAD1 transit peptide (CpSAD1tp) is indicated by uppercase, bold italics,and the remainder of the CpSAD1tp sequence located between the ATG andthe AscI site is indicated with lowercase, underlined italics. TheGarmFATA1(G108A) coding region is indicated by lowercase italics. Asequence encoding a 3×FLAG tag fused to the C-terminus ofGarmFATA1(G108A) is represented by uppercase italics, and the TGAterminator codon is indicated with uppercase, bold italics. The P.moriformis SAD2-1 3′ UTR is indicated by lowercase underlined text. TheFAD2-1 3′ genomic region is indicated by bold, lowercase text.

Nucleotide sequence of transforming DNA contained in pSZ6384SEQ ID NO: 129  gctcttcgcgaaggtcattttccagaacaacgaccatggcttgtcttagcgatcgctcgaatgactgctagtgagtcgtacgctcgacccagtcgctcgcaggagaacgcggcaactgccgagcttcggcttgccagtcgtgactcgtatgtgatcaggaatcattggcattggtagcattataattcggcttccgcgctgtttatgggcatggcaatgtctcatgcagtcgaccttagtcaaccaattctgggtggccagctccgggcgaccgggctccgtgtcgccgggcaccacctcctgccatgagtaacagggccgccctctcctcccgacgttggccaactgaataccgtgtcttggggccctacatgatgggctgcctagtcgggcgggacgcgcaactgcccgcgcaatctgggacgtggtctgaatcctccaggcgggtttccccgagaaagaaagggtgccgatttcaaagcagagccatgtgccgggccctgtggcctgtgttggcgcctatgtagtcaccccccctcacccaattgtcgccagtttgcgcaatccataaactcaaaactgcagcttctgagctgcgctgttcaa

ctgatgtccgtggtctgcaacaacaagaaccactccgcccgccccaagctgcccaactcctccctgctgcccggcttcgacgtggtggtccaggccgcggccacccgcttcaagaaggagacgacgaccacccgcgccacgctgacgttcgacccccccacgaccaactccgagcgcgccaagcagcgcaagcacaccatcgacccctcctcccccgacttccagcccatcccctccttcgaggagtgcttccccaagtccacgaaggagcacaaggaggtggtgcacgaggagtccggccacgtcctgaaggtgcccttccgccgcgtgcacctgtccggcggcgagcccgccttcgacaactacgacacgtccggcccccagaacgtcaacgcccacatcggcctggcgaagctgcgcaaggagtggatcgaccgccgcgagaagctgggcacgccccgctacacgcagatgtactacgcgaagcagggcatcatcacggaggagatgctgtactgcgcgacgcgcgagaagctggaccccgagttcgtccgctccgaggtcgcgcggggccgcgccatcatcccctccaacaagaagcacctggagctggagcccatgatcgtgggccgcaagttcctggtgaaggtgaacgcgaacatcggcaactccgccgtggcctcctccatcgaggaggaggtctacaaggtgcagtgggccaccatgtggggcgccgacaccatcatggacctgtccacgggccgccacatccacgagacgcgcgagtggatcctgcgcaactccgcggtccccgtgggcaccgtccccatctaccaggcgctggagaaggtggacggcatcgcggagaacctgaactgggaggtgttccgcgagacgctgatcgagcaggccgagcagggcgtggactacttcacgatccacgcgggcgtgctgctgcgctacatccccctgaccgccaagcgcctgacgggcatcgtgtcccgcggcggctccatccacgcgaagtggtgcctggcctaccacaaggagaacttcgcctacgagcactgggacgacatcctggacatctgcaaccagtacgacgtcgccctgtccatcggcgacggcctgcgccccggctccatctacgacgccaacgacacggcccagttcgccgagctgctgacccagggcgagctgacgcgccgcgcgtgggagaaggacgtgcaggtgatgaacgagggccccggccacgtgcccatgcacaagatccccgagaacatgcagaagcagctggagtggtgcaacgaggcgcccttctacaccctgggccccctgacgaccgacatcgcgcccggctacgaccacatcacctccgccatcggcgcggccaacatcggcgccctgggcaccgccctgctgtgctacgtgacgcccaaggagcacctgggcctgcccaaccgcgacgacgtgaaggcgggcgtcatcgcctacaagatcgccgcccacgcggccgacctggccaagcagcacccccacgcccaggcgtgggacgacgcgctgtccaaggcgcgcttcgagttccgctggatggaccagttcgcgctgtccctggaccccatgacggcgatgtccttccacgacgagacgctgcccgcggacggcgcgaaggtcgcccacttctgctccatgtgcggccccaagttctgctccatgaagatcacggaggacatccgcaagtacgccgaggagaacggctacggctccgccgaggaggccatccgccagggcatggacgccatgtccgaggagttcaacatcgccaagaagacgatctccggcgagcagcacggcgaggtcggcggcgagatctacctgcccgagtcctacgtc

cgcacgcatccaacgaccgtatacgcatcgtccaatgaccgtcggtgtcctctctgcctccgttttgtgagatgtctcaggcttggtgcatcctcgggtggccagccacgttgcgcgtcgtgctgcttgcctctcttgcgcctctgtggtactggaaaatatcatcgaggcccgtttttttgctcccatttcctttccgctacatcttgaaagcaaacgacaaacgaagcagcaagcaaagagcacgaggacggtgaacaagtctgtcacctgtatacatctatttccccgcgggtgcacctactctctctcctgccccggcagagtcagctgccttacgtgacggatcccgcgtctcgaacagagcgcgcagaggaacgctgaaggtctcgcctctgtcgcacctcagcgcggcatacaccacaataaccacctgacgaatgcgcttggttcttcgtccattagcgaagcgtccggttcacacacgtgccacgttggcgaggtggcaggtgacaatgatcggtgga

gaggagcgcaaggccaccggctaccgcgagactccggccgccacgagacccctccaacaccatgcacgccctgctggccatgggcatctggctgggcgccatccacttcaacgccctgctgctgctgactccttcctgacctgcccttctccaagttcctggtggtgttcggcctgctgctgctgacatgatcctgcccatcgacccctactccaagttcggccgccgcctgtcccgctacatctccaagcacgcctgctcctacttccccatcaccctgcacgtggaggacatccacgccttccaccccgaccgcgcctacgtgttcggcttcgagccccactccgtgctgcccatcggcgtggtggccctggccgacctgaccggcttcatgcccctgcccaagatcaaggtgctggcctcctccgccgtgttctacacccccacctgcgccacatctggacctggctgggcctgacccccgccaccaagaagaacttctcctccctgctggacgccggctactcctgcatcctggtgcccggcggcgtgcaggagaccaccacatggagcccggctccgagatcgccttcctgcgcgcccgccgcggcttcgtgcgcatcgccatggagatgggctcccccctggtgcccgtgttctgcttcggccagtcccacgtgtacaagtggtggaagcccggcggcaagttctacctgcagttctcccgcgccatcaagttcacccccatcttcttctggggcatcttcggctcccccctgccctaccagcaccccatgcacgtggtggtgggcaagcccatcgacgtgaagaagaacccccagcccatcgtggaggaggtgatcgaggtgcacgaccgcttcgtggaggccctgcaggacctgttcgagcgccacaaggcccaggtgggc

gctggtcgtgtgatggactgttgccgccacacttgctgccttgacctgtgaatatccctgccgcttttatcaaacagcctcagtgtgtttgatcttgtgtgtacgcgcttttgcgagttgctagctgcttgtgctatttgcgaataccacccccagcatccccttccctcgtttcatatcgcttgcatcccaaccgcaacttatctacgctgtcctgctatccctcagcgctgctcctgctcctgctcactgcccctcgcacagccttggtttgggctccgcctgtattctcctggtactgcaacctgtaaaccagcactgcaatgctgatgcacgggaagtagtgggatgggaacacaaatggacttaaggatctaagtaagattcgaagcgctcgaccgtgccggacggactgcagccccatgtcgtagtgaccgccaatgtaagtgggctggcgtttccctgtacgtgagtcaacgtcactgcacgcgcaccaccctctcgaccggcaggaccaggcatcgcgagat

gcgacctgcgtcgctcggcgggctccgggccccggcgcccagcgaggcccctccccgtgcgc gggcgcgcc atccccccccgcatcatcgtggtgtcctcctcctcctccaaggtgaaccccctgaagaccgaggccgtggtgtcctccggcctggccgaccgcctgcgcctgggctccctgaccgaggacggcctgtcctacaaggagaagttcatcgtgcgctgctacgaggtgggcatcaacaagaccgccaccgtggagaccatcgccaacctgctgcaggaggtgggctgcaaccacgcccagtccgtgggctactccaccgccggcttctccaccacccccaccatgcgcaagctgcgcctgatctgggtgaccgcccgcatgcacatcgagatctacaagtaccccgcctggtccgacgtggtggagatcgagtcctggggccagggcgagggcaagatcggcacccgccgcgactggatcctgcgcgactacgccaccggccaggtgatcggccgcgccacctccaagtgggtgatgatgaaccaggacacccgccgcctgcagaaggtggacgtggacgtgcgcgacgagtacctggtgcactgcccccgcgagctgcgcctggccttccccgaggagaacaactcctccctgaagaagatctccaagctggaggacccctcccagtactccaagctgggcctggtgccccgccgcgccgacctggacatgaaccagcacgtgaacaacgtgacctacatcggctgggtgctggagtccatgccccaggagatcatcgacacccacgagctgcagaccatcaccctggactaccgccgcgagtgccagcacgacgacgtggtggactccctgacctcccccgagccctccgaggacgccgaggccgtgttcaaccacaacggcaccaacggctccgccaacgtgtccgccaacgaccacggctgccgcaacttcctgcacctgctgcgcctgtccggcaacggcctggagatcaaccgcggccgcaccgagtggcgcaagaagcccacccgcATGGACTACAA

ggagcgacgagtgtgcgtgcggggctggcgggagtgggacgccctcctcgctcctctctgttctgaacggaacaatcggccaccccgcgctacgcgccacgcatcgagcaacgaagaaaaccccccgatgataggttgcggtggctgccgggatatagatccggccgcacatcaaagggcccctccgccagagaagaagctcctttcccagcagactccttctgctgccaaaacacttctctgtccacagcaacaccaaaggatgaacagatcaacttgcgtctccgcgtagcttcctcggctagcgtgcttgcaacaggtccctgcactattatcttcctgctttcctctgaattatgcggcaggcgagcgctcgctctggcgagcgctccttcgcgccgccctcgctgatcgagtgtacagtcaatgaatggtgagctcctcactcagcgcgcctgcgcggggatgcggaacgccgccgccgccttgtcttttgcacgcgcgactccgtcgcttcgcgggtggcacccccattgaaaaaaacctcaattctgtttgtggaagacacggtgtacccccaaccacccacctgcacctctattattggtattattgacgcgggagcgggcgttgtactctacaacgtagcgtctctggttttcagctggctcccaccattgtaaattcttgctaaaatagtgcgtggttatgtgagaggtatggtgtaacagggcgtcagtcatgttggttttcgtgctgatctcgggcacaaggcgtcgtcgacgtgacgtgcccgtgatgagagcaataccgcgctcaaagccgacgcatggcctttactccgcactccaaacgactgtcgctcgtatttttcggatatctattttttaagagcgagcacagcgccgggcatgggcctgaaaggcctcgcggccgtgctcgtggtgggggccgcgagcgcgtggggcatcgcggcagtgcaccaggcgcagacggaggaacgcatggtgagtgcgcatcacaagatgcatgtcttgttgtctgtactataatgctagagcatcaccaggggcttagtcatcgcacctgctttggtcattacagaaattgcacaagggcgtcctccgggatgaggagatgtaccagctcaagctggagcggcttcgagccaagcaggagcgcggcgcatgacgacctacccacatgcgaa gagc

The sequence of the pSZ6377 transforming DNA is provided below. Relevantrestriction sites in the construct are indicated in lowercase, bold andunderlined text, and are 5′-3′ BspQI, KpnI, XbaI, SnaBI, BamHI, AvrII,SpeI, AscI, ClaI, SacI and BspQ respectively. BspQI sites delimit the 5′and 3′ ends of the transforming DNA. Proceeding in the 5′ to 3′direction, bold, lowercase sequences represent FAD2-1 5′ genomic DNAthat permits targeted integration at the FAD2-1 locus via homologousrecombination. The P. moriformis LDH1 promoter driving the expression ofthe Arabidopsis thaliana THIC gene is indicated by boxed text. Theinitiator ATG and terminator TGA for AtTHIC are indicated by uppercase,bold italics while the coding region is indicated in lowercase italics.The P. moriformis HSP90 3′ UTR is indicated by lowercase underlinedtext. A spacer sequence is represented by lowercase text. The P.moriformis SAD2-1 promoter, indicated by boxed italicized text, isutilized to drive the expression of the G. mangostana FATA1 gene. Theinitiator ATG of the sequence encoding the C. protothecoides SAD1transit peptide (CpSAD1tp) is indicated by uppercase, bold italics, andthe remainder of the CpSAD1tp sequence located between the ATG and theAscI site is indicated with lowercase, underlined italics. TheGarmFATA1(G108A) coding region is indicated by lowercase italics. Asequence encoding a 3×FLAG tag fused to the C-terminus ofGarmFATA1(G108A) is represented by uppercase italics, and the TGAterminator codon is indicated with uppercase, bold italics. The P.moriformis SAD2-1 3′ UTR is indicated by lowercase underlined text. TheFAD2-1 3′ genomic region is indicated by bold, lowercase text.

Nucleotide sequence of transforming DNA contained in pSZ6377SEQ ID NO:130  gctcttcgcgaaggtcattttccagaacaacgaccatggcttgtcttagcgatcgctcgaatgactgctagtgagtcgtacgctcgacccagtcgctcgcaggagaacgcggcaactgccgagcttcggcttgccagtcgtgactcgtatgtgatcaggaatcattggcattggtagcattataattcggcttccgcgctgtttatgggcatggcaatgtctcatgcagtcgaccttagtcaaccaattctgggtggccagctccgggcgaccgggctccgtgtcgccgggcaccacctcctgccatgagtaacagggccgccctctcctcccgacgttggccaactgaataccgtgtcttggggccctacatgatgggctgcctagtcgggcgggacgcgcaactgcccgcgcaatctgggacgtggtctgaatcctccaggcgggtttccccgagaaagaaagggtgccgatttcaaagcagagccatgtgccgggccctgtggcctgtgttggcgcctatgtagtcaccccccctcacccaattgtcgccagtttgcgcaatccataaactcaaaactgcagcttctgagctgcgctgttcaa

ctgatgtccgtggtctgcaacaacaagaaccactccgcccgccccaagctgcccaactcctccctgctgcccggcttcgacgtggtggtccaggccgcggccacccgcttcaagaaggagacgacgaccacccgcgccacgctgacgttcgacccccccacgaccaactccgagcgcgccaagcagcgcaagcacaccatcgacccctcctcccccgacttccagcccatcccctccttcgaggagtgcttccccaagtccacgaaggagcacaaggaggtggtgcacgaggagtccggccacgtcctgaaggtgcccttccgccgcgtgcacctgtccggcggcgagcccgccttcgacaactacgacacgtccggcccccagaacgtcaacgcccacatcggcctggcgaagctgcgcaaggagtggatcgaccgccgcgagaagctgggcacgccccgctacacgcagatgtactacgcgaagcagggcatcatcacggaggagatgctgtactgcgcgacgcgcgagaagctggaccccgagttcgtccgctccgaggtcgcgcggggccgcgccatcatcccctccaacaagaagcacctggagctggagcccatgatcgtgggccgcaagttcctggtgaaggtgaacgcgaacatcggcaactccgccgtggcctcctccatcgaggaggaggtctacaaggtgcagtgggccaccatgtggggcgccgacaccatcatggacctgtccacgggccgccacatccacgagacgcgcgagtggatcctgcgcaactccgcggtccccgtgggcaccgtccccatctaccaggcgctggagaaggtggacggcatcgcggagaacctgaactgggaggtgttccgcgagacgctgatcgagcaggccgagcagggcgtggactacttcacgatccacgcgggcgtgctgctgcgctacatccccctgaccgccaagcgcctgacgggcatcgtgtcccgcggcggctccatccacgcgaagtggtgcctggcctaccacaaggagaacttcgcctacgagcactgggacgacatcctggacatctgcaaccagtacgacgtcgccctgtccatcggcgacggcctgcgccccggctccatctacgacgccaacgacacggcccagttcgccgagctgctgacccagggcgagctgacgcgccgcgcgtgggagaaggacgtgcaggtgatgaacgagggccccggccacgtgcccatgcacaagatccccgagaacatgcagaagcagctggagtggtgcaacgaggcgcccttctacaccctgggccccctgacgaccgacatcgcgcccggctacgaccacatcacctccgccatcggcgcggccaacatcggcgccctgggcaccgccctgctgtgctacgtgacgcccaaggagcacctgggcctgcccaaccgcgacgacgtgaaggcgggcgtcatcgcctacaagatcgccgcccacgcggccgacctggccaagcagcacccccacgcccaggcgtgggacgacgcgctgtccaaggcgcgcttcgagttccgctggatggaccagttcgcgctgtccctggaccccatgacggcgatgtccttccacgacgagacgctgcccgcggacggcgcgaaggtcgcccacttctgctccatgtgcggccccaagttctgctccatgaagatcacggaggacatccgcaagtacgccgaggagaacggctacggctccgccgaggaggccatccgccagggcatggacgccatgtccgaggagttcaacatcgccaagaagacgatctccggcgagcagcacggcgaggtcggcggcgagatctacctgcccgagtcctacgtc

cgcacgcatccaacgaccgtatacgcatcgtccaatgaccgtcggtgtcctctctgcctccgttttgtgagatgtctcaggcttggtgcatcctcgggtggccagccacgttgcgcgtcgtgctgcttgcctctcttgcgcctctgtggtactggaaaatatcatcgaggcccgtttttttgctcccatttcctttccgctacatcttgaaagcaaacgacaaacgaagcagcaagcaaagagcacgaggacggtgaacaagtctgtcacctgtatacatctatttccccgcgggtgcacctactctctctcctgccccggcagagtcagctgccttacgtgacggatcccgcgtctcgaacagagcgcgcagaggaacgctgaaggtctcgcctctgtcgcacctcagcgcggcatacaccacaataaccacctgacgaatgcgcttggttcttcgtccattagcgaagcgtccggttcacacacgtgccacgttggcgaggtggcaggtgacaatgatcggtgga

ccgcatccactttctcggcgttcaatgcccgctgcggcgacctgcgtcgctcggcgggctccgggccccggcgcccagcgaggcccctccccgtgcgcgggcgcgccatccccccccgcatcatcgtggtgtcctcctcctcctccaaggtgaaccccctgaagaccgaggccgtggtgtcctccggcctggccgaccgcctgcgcctgggctccctgaccgaggacggcctgtcctacaaggagaagttcatcgtgcgctgctacgaggtgggcatcaacaagaccgccaccgtggagaccatcgccaacctgctgcaggaggtgggctgcaaccacgcccagtccgtgggctactccaccgccggcttctccaccacccccaccatgcgcaagctgcgcctgatctgggtgaccgcccgcatgcacatcgagatctacaagtaccccgcctggtccgacgtggtggagatcgagtcctggggccagggcgagggcaagatcggcacccgccgcgactggatcctgcgcgactacgccaccggccaggtgatcggccgcgccacctccaagtgggtgatgatgaaccaggacacccgccgcctgcagaaggtggacgtggacgtgcgcgacgagtacctggtgcactgcccccgcgagctgcgcctggccttccccgaggagaacaactcctccctgaagaagatctccaagctggaggacccctcccagtactccaagctgggcctggtgccccgccgcgccgacctggacatgaaccagcacgtgaacaacgtgacctacatcggctgggtgctggagtccatgccccaggagatcatcgacacccacgagctgcagaccatcaccctggactaccgccgcgagtgccagcacgacgacgtggtggactccctgacctcccccgagccctccgaggacgccgaggccgtgttcaaccacaacggcaccaacggctccgccaacgtgtccgccaacgaccacggctgccgcaacttcctgcacctgctgcgcctgtccggcaacggcctggagatcaaccgcggccgcaccgagtggcgcaagaagcccacccgcATGGACTACAAGGACCACGACGGCGACTACAAGGACCACGACATCGACTACA

ctctctgttctgaacggaacaatcggccaccccgcgctacgcgccacgcatcgagcaacgaagaaaaccccccgatgataggttgcggtggctgccgggatatagatccggccgcacatcaaagggcccctccgccagagaagaagctcctttcccagcagactccttctgctgccaaaacacttctctgtccacagcaacaccaaaggatgaacagatcaacttgcgtctccgcgtagcttcctcggctagcgtgcttgcaacaggtccctgcactattatcttcctgctttcctctgaattatgcggcaggcgagcgctcgctctggcgagcgctccttcgcgccgccctcgctgatcgagtgtacagtcaatgaatggtgagctcctcactcagcgcgcctgcgcggggatgcggaacgccgccgccgccttgtcttttgcacgcgcgactccgtcgcttcgcgggtggcacccccattgaaaaaaacctcaattctgtttgtggaagacacggtgtacccccaaccacccacctgcacctctattattggtattattgacgcgggagcgggcgttgtactctacaacgtagcgtctctggttttcagctggctcccaccattgtaaattcttgctaaaatagtgcgtggttatgtgagaggtatggtgtaacagggcgtcagtcatgttggttttcgtgctgatctcgggcacaaggcgtcgtcgacgtgacgtgcccgtgatgagagcaataccgcgctcaaagccgacgcatggcctttactccgcactccaaacgactgtcgctcgtatttttcggatatctattttttaagagcgagcacagcgccgggcatgggcctgaaaggcctcgcggccgtgctcgtggtgggggccgcgagcgcgtggggcatcgcggcagtgcaccaggcgcagacggaggaacgcatggtgagtgcgcatcacaagatgcatgtcttgttgtctgtactataatgctagagcatcaccaggggcttagtcatcgcacctgctttggtcattacagaaattgcacaagggcgtcctccgggatgaggagatgtaccagctcaagctggagcggcttcgagccaagcaggagcgcggcgcatgacgacctacccacatgc gaagagc

Constructs pSZ6383, pSZ6384 and pSZ6377 were transformed into S8813.Primary transformants were clonally purified and screened under standardlipid production conditions at pH 5. Integration of pSZ6383 or pSZ6384at the FAD2-1 locus was verified by DNA blot analysis. The fatty acidprofiles, sn-2 profiles and lipid titers of lead strains were assayed in50-mL shake flasks (Table 23). FAD2-1 ablation reduced C18:2 to <1% inmost strains. Expression of a second copy of GarmFATA1(G108A) andTcDGAT1 (S8990, S8992, S8998 & S8999), or TcDGAT2 (S8994, S9000 & S9047)elevated C18:0 to >56%. The D5393-28 strain, expressing a second copy ofGarmFATA1(G108A) without either of the cocoa DGAT genes (pSZ6377) had asimilar fatty acid profile, but lower lipid titer. As shown in Table 23,as compared to strain S8813, for strains expressing either TcDGAT1 orTcDGAT2, C16:0 increased from 3.2% to 3.7%-4.0%, C18:0 increased from45.8% to about 56%, C18:2 decreased from 1.4% to about 1.0%.

TABLE 23 Fatty acid profiles of FAD2-1 ablation strains. Strain S8813D5393-28 S8990 S8992 S8998 S8999 S8994 S9000 S9047 C12:0 0.1 0.2 0.2 0.20.1 0.2 0.1 0.1 0.2 C14:0 0.4 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 C16:0 3.23.8 3.7 3.8 3.9 4.0 3.7 3.8 3.5 C16:1 cis-7 0.1 0.1 0.1 0.1 0.1 0.1 0.10.1 0.1 C16:1 cis-9 0.0 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 C17:0 0.1 0.20.2 0.1 0.2 0.1 0.2 0.2 0.2 C18:0 45.8 56.0 56.6 56.0 56.2 56.0 56.356.4 56.5 C18:1 45.9 35.8 35.4 35.9 35.7 35.5 35.9 35.7 35.9 C18:2 1.41.0 0.9 1.0 0.9 1.1 0.9 0.9 0.8 C18:3 α 0.3 0.3 0.3 0.2 0.3 0.2 0.2 0.30.3 C20:0 2.0 1.6 1.6 1.5 1.6 1.5 1.5 1.5 1.5 C22:0 0.2 0.2 0.2 0.2 0.20.2 0.2 0.2 0.2 C24:0 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 saturates 52.162.6 63.1 62.6 62.9 62.8 62.8 62.9 62.7

Liquid chromatography and mass spectrometry were used to analyze the TAGcomposition of final strains. The strains accumulated 68-71% SOS, withtrisaturates ranging from 2.5-2.8%. The D5393-28 strain, expressing asecond copy of GarmFATA1(G108A) without either of the cocoa DGAT geneshad similar SOS content but slightly higher trisaturates. The TAGcomposition of a typical Shea stearin and a sample of Kokum butter areshown for comparison

TABLE 24 LC/MS TAG profiles of FAD2-1 ablation strains. Shea KokumStrain D5393-28 S8990 S8992 S8998 S8999 S8994 S9000 S9047 stearin butterOOL 0.4 LLS 0.2 POL 0.3 OOO 1.3 1.7 SOL 1.0 0.4 LaOS + MOP 0.2 0.3 0.30.2 0.3 0.3 0.4 0.2 OOP 0.5 0.2 0.3 0.2 0.2 0.4 0.3 0.2 0.8 0.7 PLS(+SLnS) 0.6 0.7 0.7 0.7 0.7 0.6 0.6 0.4 0.6 0.3 POP (+MOS) 1.1 1.0 1.01.1 1.1 1.0 1.2 0.8 0.7 0.4 OOS 10.5 10.3 11.3 11.0 11.0 10.9 10.1 10.66.4 11.8 SLS (+PLA) 1.9 1.7 2.0 1.6 2.1 1.8 1.9 1.5 5.5 1.4 POS 8.4 8.58.4 8.7 8.9 8.4 10.0 7.7 6.3 4.8 MaOS 0.3 SOG 0.4 0.5 0.5 0.6 0.3 0.50.4 0.5 OOA 0.5 0.3 0.4 0.4 0.4 0.4 0.4 0.4 0.2 0.2 SOS (+POA) 68.4 69.768.7 69.1 68.3 69.4 68.0 71.4 69.7 76.6 SSP (+MSA) 0.5 0.5 0.5 0.4 0.50.5 0.5 0.4 0.2 SOA + POB 3.9 3.8 3.5 3.6 3.4 3.5 3.5 3.4 4.0 1.0 SSS(+PSA) 2.6 2.3 2.2 2.1 2.3 2.2 2.3 2.1 2.0 0.5 SOB + LgOP + AOA 0.4 0.20.2 0.3 0.3 0.3 0.3 0.3 0.4 SSA (+PBS) 0.2 SOLg (+POHx) 0.3 SUM (area %)99.8 99.9 99.8 99.9 99.8 99.9 100.0 99.9 100.0 100.0 Sat-Sat-Sat 3.1 2.82.7 2.5 2.7 2.7 2.8 2.5 2.4 0.5 Sat-U-Sat 84.9 85.9 84.7 85.3 85.1 85.086.0 85.8 87.5 84.7 Sat-O-Sat 82.4 83.5 82.0 82.9 82.3 82.6 83.4 83.981.4 83.1 Sat-L-Sat 2.5 2.4 2.6 2.3 2.8 2.4 2.6 1.9 6.1 1.6 U-U-U/Sat11.8 11.3 12.4 12.2 12.0 12.2 11.3 11.7 10.6 14.8 La = laurate (C12:0),M = myristate (C14:0), P = palmitate (C16:0), Ma = margarate (C17:0), S= stearate (C18:0), O = oleate (C18:1), L = linoleate (C18:2), Ln =α-linolenate (C18:3 α), A = arachidate (C20:0), G = (C20:1), B =behenate (C22:0), Lg = lignocerate (C24:0), Hx = hexacosanoate (C26:0).Sat = saturated, U = unsaturated

Example 8 Variant Brassica napus Thioeserase

In this example, we demonstrate the modification of the enzymespecificity of a FATA thioesterase originally isolated from Brassicanapus (BnOTE, accession CAA52070), by site directed mutagenesistargeting two amino acids positions D124 and D209).

To determine the impact of each amino acid substitution on the enzymespecificity of the BnOTE, the wild-type and the mutant BnOTE genes werecloned into a vector enabling expression and expressed in P. moriformisstrain S8588. Strain S8588 is a strain in which the endogenous FATA1allele has been disrupted and expresses a Prototheca moriformis KASIIgene and sucrose invertase. Recombinant strains with FATA1 disruptionand co-expression of P. moriformis KASII and invertase were previouslydisclosed in co-owned applications WO2012/106560 and WO2013/15898,herein incorporated by reference.

Strains that express wild type or mutant BnOTE enzymes, constructspSZ6315, pSZ6316, pSZ6317, or pSZ6318 were expressed in S8588. In theseconstructs, the Saccharomyces carlsbergensis MEL1 gene (Accession no:AAA34770) was utilized as the selectable marker to introduce thewild-type and mutant BnOTE genes into the FAD2-2 locus of P. moriformisstrain S8588 by homologous recombination using previously describedtransformation methods (biolistics). The constructs that have beenexpressed in S8588 are listed in Table 25.

TABLE 25 DNA lot# and plasmid ID of DNA constructs that expressingwild-type and mutant BnOTE genes DNA Solazynne Lot# Plasmid ConstructD5309 pSZ6315 FAD2-2::PmHXT1-ScarMEL1-PmPGK:PmSAD2-2V3-CpSADtp-BnOTE-PmSAD2-1 utr::FAD2-2 D5310 pSZ6316FAD2-2::PmHXT1-ScarMEL1-PmPGK:PmSAD2-2 V3-CpSADtp-BnOTE(D124A)-PmSAD2-1utr::FAD2-2 D5311 pSZ6317 FAD2-2::PmHXT1-ScarMEL1-PmPGK:PmSAD2-2V3-CpSADtp-BnOTE(D209A)-PmSAD2-1 utr::FAD2-2 D5312 pSZ6318FAD2-2::PmHXT1-ScarMEL1-PmPGK:PmSAD2-2 V3-CpSADtp-BnOTE(D124A, D209A)-PmSAD2-1 utr::FAD2-2

pSZ6315

The construct psZ6315 can be written asFAD2-2::PmHXT1-ScarMEL1-PmPGK:PmSAD2-2 V3-CpSADtp-BnOTE-PmSAD2-1utr::FAD2-2. The sequence of the pSZ6315 transforming DNA is providedbelow. Relevant restriction sites in pSZ6315 are indicated in lowercase,bold and underlining and are 5′-3′ SgrAI, Kpn I, SnaBI, AvrII, SpeI,AscI, ClaI, Sac I, SbfI, respectively. SgrAI and SbfI sites delimit the5′ and 3′ ends of the transforming DNA. Bold, lowercase sequencesrepresent FAD2-2 genomic DNA that permit targeted integration at FAD2-2locus via homologous recombination. Proceeding in the 5′ to 3′direction, the P. moriformis HXT1 promoter driving the expression of theSaccharomyces carlsbergensis MEL1 gene is indicated by boxed text. Theinitiator ATG and terminator TGA for MEL1 gene are indicated byuppercase, bold italics while the coding region is indicated inlowercase italics. The P. moriformis PGK 3′ UTR is indicated bylowercase underlined text followed by the P. moriformis SAD2-2 V3promoter, indicated by boxed italics text. The Initiator ATG andterminator TGA codons of the wild-type BnOTE are indicated by uppercase,bold italics, while the remainder of the coding region is indicated bybold italics in lower case. The three-nucleotide codon corresponding tothe target amino acids, D124 and D209, are in lower case, italicized,bolded and wave underlined. The P. moriformis SAD2-1 3′UTR is againindicated by lowercase underlined text followed by the FAD2-2 genomicregion indicated by bold, lowercase text.

Nucleotide sequence of transforming DNA contained in pSZ6315SEQ ID NO: 131  caccggcgcgctgcttcgcgtgccgggtgcagcaatcagatccaagtctgacgacttgcgcgcacgcgccggatccttcaattccaaagtgtcgtccgcgtgcgcttcttcgccttcgtcctcttgaacatccagcgacgcaagcgcagggcgctgggcggctggcgtcccgaaccggcctcggcgcacgcggctgaaattgccgatgtcggcaatgtagtgccgctccgcccacctctcaattaagtttttcagcgcgtggttgggaatgatctgcgctcatggggcgaaagaaggggttcagaggtgctttattgttactcgactgggcgtaccagcattcgtgcatgactgattatacatacaaaagtacagctcgcttcaatgccctgcgattcctactcccgagcgagcactcctctcaccgtcgggttgcttcccacgaccacgccggtaagagggtctgtggcctcgcgcccctcgcgagcgcatattccagccacgtctgtatgattttgcgctcatacgtctggcccgtcgaccccaaaatgacgggatcctgcataatatcgcccgaaatgggatccaggcattcgtcaggaggcgtcagccccgcgggagatgccggtcccgccgcattggaaaggtgtagagggggt

gcctgggcctgacgccccagatgggctgggacaactggaacacgttcgcctgcgacgtctccgagcagctgctgctggacacggccgaccgcatctccgacctgggcctgaaggacatgggctacaagtacatcatcctggacgactgctggtcctccggccgcgactccgacggcttcctggtcgccgacgagcagaagttccccaacggcatgggccacgtcgccgaccacctgcacaacaactccttcctgttcggcatgtactcctccgcgggcgagtacacgtgcgccggctaccccggctccctgggccgcgaggaggaggacgcccagttcttcgcgaacaaccgcgtggactacctgaagtacgacaactgctacaacaagggccagttcggcacgcccgagatctcctaccaccgctacaaggccatgtccgacgccctgaacaagacgggccgccccatcttctactccctgtgcaactggggccaggacctgaccttctactggggctccggcatcgcgaactcctggcgcatgtccggcgacgtcacggcggagttcacgcgccccgactcccgctgcccctgcgacggcgacgagtacgactgcaagtacgccggcttccactgctccatcatgaacatcctgaacaaggccgcccccatgggccagaacgcgggcgtcggcggctggaacgacctggacaacctggaggtcggcgtcggcaacctgacggacgacgaggagaaggcgcacttctccatgtgggccatggtgaagtcccccctgatcatcggcgcgaacgtgaacaacctgaaggcctcctcctactccatctactcccaggcgtccgtcatcgccatcaaccaggactccaacggcatccccgccacgcgcgtctggcgctactacgtgtccgacacggacgagtacggccagggcgagatccagatgtggtccggccccctggacaacggcgaccaggtcgtggcgctgctgaacggcggctccgtgtcccgccccatgaacacgaccctggaggagatcttcttcgactccaacctgggctccaagaagctgacctccacctgggacatctacgacctgtgggcgaaccgcgtcgacaactccacggcgtccgccatcctgggccgcaacaagaccgccaccggcatcctgtacaacgccaccgagcagtcctacaaggacggcctgtccaagaacgacacccgcctgttcggccagaagatcggctccctgtc

accggcgctgatgtggcgcggacgccgtcgtactctttcagactttactcttgaggaattgaacctttctcgcttgctggcatgtaaacattggcgcaattaattgtgtgatgaagaaagggtggcacaagatggatcgcgaatgtacgagatcgacaacgatggtgattgttatgaggggccaaacctggctcaatcttgtcgcatgtccggcgcaatgtgatccagcggcgtgactctcgcaacctggtagtgtgtgcgcaccgggtcgctttgattaaaactgatcgcattgccatcccgtcaactcacaagcctactctagctcccattgcgcactcgggcgcccggctcgatcaatgttctgagcggagggcgaagcgtcaggaaatcgtctcggcagctggaagcgcatggaatgcggagcggagatcgaatcaggatcccgcgtctcgaacagagcgcgcagaggaacgctgaaggtctcgcctctgtcgcacctcagcgcggcatacaccacaataaccacctgacgaatgcgcttggttcttcgtccattagcgaagcgt

tcgctcctctctgttctgaacggaacaatcggccaccccgcgctacgcgccacgcatcgagcaacgaagaaaaccccccgatgataggttgcggtggctgccgggatatagatccggccgcacatcaaagggcccctccgccagagaagaagctcctttcccagcagactccttctgctgccaaaacacttctctgtccacagcaacaccaaaggatgaacagatcaacttgcgtctccgcgtagcttcctcggctagcgtgcttgcaacaggtccctgcactattatcttcctgctttcctctgaattatgcggcaggcgagcgctcgctctggcgagcgctccttcgcgccgccctcgctgatcgagtgtacagtcaatgaatggtgagctccgcgcctgcgcgaggacgcagaacaacgctgccgccgtgtcttttgcacgcgcgactccggcgcttcgctggtggcacccccataaagaaaccctcaattctgtttgtggaagacacggtgtacccccacccacccacctgcacctctattattggtattattgacgcgggagtgggcgttgtaccctacaacgtagcttctctagttttcagctggctcccaccattgtaaattcatgctagaatagtgcgtggttatgtgagaggtatagtgtgtctgagcagacggggcgggatgcatgtcgtggtggtgatctttggctcaaggcgtcgtcgacgtgacgtgcccgatcatgagagcaataccgcgctcaaagccgacgcatagcctttactccgcaatccaaacgactgtcgctcgtattifttggatatctattttaaagagcgagcacagcgccgggcatgggcctgaaaggcctcgcggccgtgctcgtggtgggggccgcgagcgcgtggggcatcgcggcagtgcaccaggcgcagacggaggaacgcatggtgcgtgcgcaatataagatacatgtattgttgt cctgcaggNucleotide sequence of BnOTE (D124A) in pSZ6316 SEQ ID NO: 132 

The sequence of the pSZ6317 transforming DNA is same as pSZ6315 exceptthe D209A point mutation, the BnOTE D209A DNA sequence is providedbelow. The three-nucleotide codon corresponding to the target two aminoacids, D124 and D209, are in lower case, italicized, bolded and waveunderlined. pSZ6317 is written as FAD2-2::PmHXT1-ScarMEL1-PmPGK:PmSAD2-2V3-CpSADtp-BnOTE (D209A)-PmSAD2-1 utr::FAD2-2

SEQ ID NO: 133 Nucleotide sequence of BnOTE (D209A) in pSZ6317:

atggactacaaggaccacgacggcgactacaaggaccacgacatcgactacaagg acgacgacgacaag

The sequence of the pSZ6318 transforming DNA is same as pSZ6315 excepttwo point mutations, D124A and D209A, the BnOTE (D124A, D209A) DNAsequence is provided below. The three-nucleotide codon corresponding tothe target two amino acids, D124 and D209, are in lower case,italicized, bolded and wave underlined. pSZ6318 is written asFAD2-2::PmHXT1-ScarMEL1-PmPGK:PmSAD2-2 V3-CpSADtp-BnOTE (D124A,D209A)-PmSAD2-1 utr::FAD2-2

SEQ ID NO: 134 Nucleotide sequence of BnOTE (D124A, D209A) in pSZ6318

atggactacaaggaccacgacggcgactacaaggaccacgacatcgactacaagg acgacgacgacaag

The DNA constructs containing the wild-type and mutant BnOTE genes weretransformed into the parental strain S8588. Primary transformants wereclonally purified and grown under standard lipid production conditionsat pH5.0. The resulting profiles from representative clones arising fromtransformations with pSZ6315, pSZ6316, pSZ6317, and pSZ6318 into S8588are shown in Table 26. The parental strain S8588 produces 5.4% C18:0,when transformed with the DNA cassette expressing wild-type BnOTE, thetransgenic lines produce ˜11% C18:0. The BnOTE mutant (D124A) increasedthe amount of C18:0 by at least 2 fold compared to the wild-typeprotein. In contrast, the BnOTE D209A mutation appears to have no impacton the enzyme activity/specificity of the BnOTE thioesterase. Finally,expression of the BnOTE (D124A, D209A) resulted in very similar fattyacid profile to what we observed in the transformants from S8588expressing BnOTE (D124A), again indicating that D209A has no significantimpact on the enzyme activity.

TABLE 26 Fatty acid profiles in S8588 and derivative transgenic linestransformed with wild-type and mutant BnOTE genes Fatty Acid Area %Transforming DNA Sample ID C16:0 C18:0 C18:1 C18:2 pH5; S8588 (parentalstrain) 3.00 5.43 81.75 6.47 D5309, pSZ6315; pH5; S8588, D5309-6; 3.8611.68 76.51 5.06 wild-type BnOTE pH5; S8588, D5309-2; 3.50 11.00 77.804.95 pH5; S8588, D5309-9 ; 3.51 10.72 78.03 5.00 pH5; S8588, D5309-10;3.55 10.69 78.06 4.96 pH5; S8588, D5309-11; 3.61 10.69 78.05 4.95 D5310,pSZ6316, pH5; S8588, D5310-6; 4.27 31.55 55.31 5.30 BnOTE (D124A) pH5;S8588, D5310-1; 4.53 30.85 54.71 6.03 pH5; S8588, D5310-5; 5.21 20.7565.43 5.02 pH5; S8588, D5310-10; 4.99 19.18 67.75 5.00 pH5; S8588,D5310-2; 4.90 18.92 68.17 4.98 D5311, pSZ6317, pH5; S8588, D5311-3; 3.5011.90 76.95 4.98 BnOTE (D209A) pH5; S8588, D5311-4; 3.63 11.35 77.444.94 pH5; S8588, D5311-14; 3.47 11.23 77.68 4.98 pH5; S8588, D5311-10;3.60 11.20 77.53 5.00 pH5; S8588, D5311-12; 3.53 11.12 77.59 5.09 D5312,pSZ6318, pH5; S8588, D5312-20; 4.79 37.97 47.74 6.01 BnOTE (D124A, pH5;S8588, D5312-40; 5.97 22.94 62.20 5.11 D209A) pH5; S8588, D5312-39; 6.0722.75 62.24 5.17 pH5; S8588, D5312-16; 5.25 18.81 67.36 5.09 pH5; S8588,D5312-26; 4.93 18.70 68.37 4.96

Example 9 Variant Garcinia mangostana Thioeserase

In this example, we demonstrate the ability to modify the activity andspecificity of a FATA thioesterase originally isolated from Garciniamangostana (GmFATA, accession 004792), using site directed mutagenesistargeting six amino acid positions within the enzyme and variouscombinations thereof. Facciotti et al (NatBiotech 1999) had previouslyaltered three of the amino acids (G108, S111, V193). The remaining threeamino acids targeted are L91, G96, and T156.

To test the impact of each mutation on the activity of the GmFATA, thewild-type and mutant genes were cloned into a vector enabling expressionwithin the P. moriformis strain S3150. Table 27 summarizes the resultsfrom a three day lipid profile screen comparing the wild-type GmFATAwith the 14 mutants. Three GmFATA mutants (DNA lot numbers D3998, D4000,D4003) increased the amount of C18:0 by at least 1.5 fold compared tothe wild-type protein (DNA lot number D3997). D3998 and D4003 weremutations that had been described by Facciotti et al (NatBiotech 1999)as substitutions that increased the activity of the GmFATA. Strain S3150expressing the mutations contained in DNA lot number D4000 was based onresearch at Solazyme which demonstrated this position influenced theactivity of the FATB thioesterases. All of the constructs were codonoptimized to reflect UTEX 1435 codon usage. Non-mutated GmFATA increasesthe fatty acid content of C18:0 and decreases the fatty acid content ofC18:1 and C18:2. As can be seen in Table 27 the G90A mutant GmFATAincreases the fatty acid content of C18:0 and decreases the fatty acidcontent of C18:1 and C18:2 when compared to the wild-type GmFATA.

TABLE 27 Algal Strain DNA # GmFATA C14:0 C16:0 C18:0 C18:1 C18:2 P.S3150 1.63 29.82 3.08 55.95 7.22 moriformis D3997 Wild-Type 1.79 29.287.32 52.88 6.21 S3150 pSZ5083 GmFATA D3998 S111A, 1.84 28.88 11.19 49.086.21 pSZ5084 V193A D3999 S111V, 1.73 29.92 3.23 56.48 6.46 pSZ5085 V193AD4000 G96A 1.76 30.19 12.66 45.99 6.01 pSZ5086 D4001 G96T 1.82 30.603.58 55.50 6.28 pSZ5087 D4002 G96V 1.78 29.35 3.45 56.77 6.43 pSZ5088D4003 G108A 1.77 29.06 12.31 47.86 6.08 pSZ5089 D4007 G108V 1.81 28.785.71 55.05 6.26 pSZ5093 D4004 L91F 1.76 29.60 6.97 53.04 6.13 pSZ5090D4005 L91K 1.87 28.89 4.38 56.24 6.35 pSZ5091 D4006 L91S 1.85 28.06 4.8156.45 6.47 pSZ5092 D4008 T156F 1.81 28.71 3.65 57.35 6.31 pSZ5094 D4009T156A 1.72 29.66 5.44 54.54 6.26 pSZ5095 D4010 T156K 1.73 29.95 3.1756.86 6.21 pSZ5096 D4011 T156V 1.80 29.17 4.97 55.44 6.27 pSZ5097

Nucleotide sequence of the GmFATA wild-type parental gene expressionvector is shown below (D3997, pSZ5083). The plasmid pSZ5083 can bewritten as THI4a::CrTUB2-NeoR-PmPGH:PmSAD2-2Ver3-CpSAD1tp_GarmFATA1FLAG-CvNR::THI4a. The 5′ and 3′ homology arms enabling targetedintegration into the Thi4 locus are noted with lowercase; the CrTUB2promoter is noted in uppercase italic which drives expression of theneomycin selection marker noted with lowercase italic followed by thePmPGH 3′UTR terminator highlighted in uppercase. The PmSAD2-1 promoter(noted in bold text) drives the expression of the GmFATA gene (notedwith lowercase bold text) and is terminated with the CvNR 3′UTR noted inunderlined, lower case bold. Restriction cloning sites and spacer DNAfragments are noted as underlined, uppercase plain lettering. Thenucleotide sequence for all of the GmFATA constructs disclosed in thisexample is identical to that of pSZ5083 with the exception of theencoded GmFATA. The promoter, 3′UTR, selection marker and targeting armsare the same as described for pSZ5083. The individual GmFATA mutantsequences are shown below. The amino acid sequence of the unmutagenizedGmFATA is showing in FIG. 1. The amino acid sequences of the alteredGmFATA proteins are shown below.

SEQ ID NO: 135 pSZ5083ccctcaactgcgacgctgggaaccttctccgggcaggcgatgtgcgtgggtttgcctccttggcacggctctacaccgtcgagtacgccatgaggcggtgatggctgtgtcggttgccacttcgtccagagacggcaagtcgtccatcctctgcgtgtgtggcgcgacgctgcagcagtccctctgcagcagatgagcgtgactttggccatttcacgcactcgagtgtacacaatccatttttcttaaagcaaatgactgctgattgaccagatactgtaacgctgatttcgctccagatcgcacagatagcgaccatgttgctgcgtctgaaaatctggattccgaattcgaccctggcgctccatccatgcaacagatggcgacacttgttacaattcctgtcacccatcggcatggagcaggtccacttagattcccgatcacccacgcacatctcgctaatagtcattcgttcgtgtcttcgatcaatctcaagtgagtgtgcatggatcttggttgacgatgcggtatgggtttgcgccgctggctgcagggtctgcccaaggcaagctaacccagctcctctccccgacaatactctcgcaggcaaagccggtcacttgccttccagattgccaataaactcaattatggcctctgtcatgccatccatgggtctgatgaatggtcacgctcgtgtcctgaccgttccccagcctctggcgtcccctgccccgcccaccagcccacgccgcgcggcagtcgctgccaaggctgtctcggaGGTACC CTTTCTTGCGCTATGACACTTCCAGCAAAAGGTAGGGCGGGCTGCGAGACGGCTTCCCGGCGCTGCATGCAACACCGATGATGCTTCGACCCCCCGAAGCTCCTTCGGGGCTGCATGGGCGCTCCGATGCCGCTCCAGGGCGAGCGCTGTTTAAATAGCCAGGCCCCCGATTGCAAAGACATTATAGCGAGCTACCAAAGCCATATTCAAACACCTAGATCACTACCACTTCTACACAGGCCACTCGAGCTTGTGATCGCACTCCGCTAAGGGGGCGCCTCTTCCTCTTCGTTTCAGTCACAACCCGCAAAC TCTAGAATATC Aatgatcgagcaggacggcctccacgccggctcccccgccgcctgggtggagcgcctgttcggctacgactgggcccagcagaccatcggctgctccgacgccgccgtgttccgcctgtccgcccagggccgccccgtgctgttcgtgaagaccgacctgtccggcgccctgaacgagctgcaggacgaggccgcccgcctgtcctggctggccaccaccggcgtgccctgcgccgccgtgctggacgtggtgaccgaggccggccgcgactggctgctgctgggcgaggtgcccggccaggacctgctgtcctcccacctggcccccgccgagaaggtgtccatcatggccgacgccatgcgccgcctgcacaccctggaccccgccacctgccccttcgaccaccaggccaagcaccgcatcgagcgcgcccgcacccgcatggaggccggcctggtggaccaggacgacctggacgaggagcaccagggcctggcccccgccgagctgttcgcccgcctgaaggcccgcatgcccgacggcgaggacctggtggtgacccacggcgacgcctgcctgcccaacatcatggtggagaacggccgcttctccggcttcatcgactgcggccgcctgggcgtggccgaccgctaccaggacatcgccctggccacccgcgacatcgccgaggagctgggcggcgagtgggccgaccgcttcctggtgctgtacggcatcgccgcccccgactcccagcgcatcgccttctaccgcctgctggacgagttcttctga CAATTGACGCCCGCGCGGCGCACCTGACCTGTTCTCTCGAGGGCGCCTGTTCTGCCTTGCGAAACAAGCCCCTGGAGCATGCGTGCATGATCGTCTCTGGCGCCCCGCCGCGCGGTTTGTCGCCCTCGCGGGCGCCGCGGCCGCGGGGGCGCATTGAAATTGTTGCAAACCCCACCTGACAGATTGAGGGCCCAGGCAGGAAGGCGTTGAGATGGAGGTACAGGAGTCAAGTAACTGAAAGTTTTTATGATAACTAACAACAAAGGGTCGTTTCTGGCCAGCGAATGACAAGAACAAGATTCCACATTTCCGTGTAGAGGCTTGCCATCGAATGTGAGCGGGCGGGCCGCGGACCCGACAAAACCCTTACGACGTGGTAAGAAAAACGTGGCGGGCACTGTCCCTGTAGCCTGAAGACCAGCAGGAGACGATCGGAAGCATCACAGCACAGGATCCCGCGTCTCGAACAGAGCGCGCAGAGGAACGCTGAAGGTCTCGCCTCTGTCGCACCTCAGCGCGGCATACACCACAATAACCACCTGACGAATGCGCTTGGTTCTTCGTCCATTAGCGAAGCGTCCGGTTCACACACGTGCCACGTTGGCGAGGTGGCAGGTGACAATGATCGGTGGAGCTGATGGTCGAAACGTTCACAGCCTAGGGATATC GTGAAAACTCGCTCGACCGCCCGCGTCCCGCAGGCAGCGATGACGTGTGCGTGACCTGGGTGTTTCGTCGAAAGGCCAGCAACCCCAAATCGCAGGCGATCCGGAGATTGGGATCTGATCCGAGCTTGGACCAGATCCCCCACGATGCGGCACGGGAACTGCATCGACTCGGCGCGGAACCCAGCTTTCGTAAATGCCAGATTGGTGTCCGATACCTTGATTTGCCATCAGCGAAACAAGACTTCAGCAGCGAGCGTATTTGGCGGGCGTGCTACCAGGGTTGCATACATTGCCCATTTCTGTCTGGACCGCTTTACCGGCGCAGAGGGTGAGTTGATGGGGTTGGCAGGCATCGAAACGCGCGTGCATGGTGTGTGTGTCTGTTTTCGGCTGCACAATTTCAATAGTCGGATGGGCGACGGTAGAATTGGGTGTTGCGCTCGCGTGCATGCCTCGCCCCGTCGGGTGTCATGACCGGGACTGGAATCCCCCCTCGCGACCCTCCTGCTAACGCTCCCGACTCTCCCGCCCGCGCGCAGGATAGACTCTAGTTCAACCAATCGACA ACTAGT atggccaccgcatccactttctcggcgttcaatgcccgctgcggcgacctgcgtcgctcggcgggctccgggccccggcgcccagcgaggcccctccccgtgcgcgggcgcgccatccccccccgcatcatcgtggtgtcctcctcctcctccaaggtgaaccccctgaagaccgaggccgtggtgtcctccggcctggccgaccgcctgcgcctgggctccctgaccgaggacggcctgtcctacaaggagaagttcatcgtgcgctgctacgaggtgggcatcaacaagaccgccaccgtggagaccatcgccaacctgctgcaggaggtgggctgcaaccacgcccagtccgtgggctactccaccggcggcttctccaccacccccaccatgcgcaagctgcgcctgatctgggtgaccgcccgcatgcacatcgagatctacaagtaccccgcctggtccgacgtggtggagatcgagtcctggggccagggcgagggcaagatcggcacccgccgcgactggatcctgcgcgactacgccaccggccaggtgatcggccgcgccacctccaagtgggtgatgatgaaccaggacacccgccgcctgcagaaggtggacgtggacgtgcgcgacgagtacctggtgcactgcccccgcgagctgcgcctggccttccccgaggagaacaactcctccctgaagaagatctccaagctggaggacccctcccagtactccaagctgggcctggtgccccgccgcgccgacctggacatgaaccagcacgtgaacaacgtgacctacatcggctgggtgctggagtccatgccccaggagatcatcgacacccacgagctgcagaccatcaccctggactaccgccgcgagtgccagcacgacgacgtggtggactccctgacctcccccgagccctccgaggacgccgaggccgtgttcaaccacaacggcaccaacggctccgccaacgtgtccgccaacgaccacggctgccgcaacttcctgcacctgctgcgcctgtccggcaacggcctggagatcaaccgcggccgcaccgagtggcgcaagaagcccacccgcatggactacaaggaccacgacggcgactacaaggaccacgacatcgactacaaggacgacgacgacaagtga ATCGATgcagcagcagctcggatagtatcgacacactctggacgctggtcgtgtgatggactgttgccgccacacttgctgccttgacctgtgaatatccctgccgcttttatcaaacagcctcagtgtgtttgatcttgtgtgtacgcgcttttgcgagttgctagctgcttgtgctatttgcgaataccacccccagcatccccttccctcgtttcatatcgcttgcatcccaaccgcaacttatctacgctgtcctgctatccctcagcgctgctcctgctcctgctcactgcccctcgcacagccttggtttgggctccgcctgtattctcctggtactgcaacctgtaaaccagcactgcaatgctgatgcacgggaagtagtgggatgggaacacaaatggaAAGCTTGAGCTCcagcgccatgccacgccctttgatggcttcaagtacgattacggtgttggattgtgtgtttgttgcgtagtgtgcatggtttagaataatacacttgatttcttgctcacggcaatctcggcttgtccgcaggttcaaccccatttcggagtctcaggtcagccgcgcaatgaccagccgctacttcaaggacttgcacgacaacgccgaggtgagctatgtttaggacttgattggaaattgtcgtcgacgcatattcgcgctccgcgacagcacccaagcaaaatgtcaagtgcgttccgatttgcgtccgcaggtcgatgttgtgatcgtcggcgccggatccgccggtctgtcctgcgcttacgagctgaccaagcaccctgacgtccgggtacgcgagctgagattcgattagacataaattgaagattaaacccgtagaaaaatttgatggtcgcgaaactgtgctcgattgcaagaaattgatcgtcctccactccgcaggtcgccatcatcgagcagggcgttgctcccggcggcggcgcctggctggggggacagctgttctcggccatgtgtgtacgtagaaggatgaatttcagctggttttcgttgcacagctgtttgtgcatgatttgtttcagactattgttgaatgtttttagatttcttaggatgcatgatttgtctgcatgcgactSEQ ID NO: 136 Amino acid sequence of Gm FATA wild-type parental gene;D3997, pSZ5083. The algal transit peptide is underlined and the FLAG epitope tag isuppercase boldMATASTFSAFNARCGDLRRSAGSGPRRPARPLPVRGRAIPPRIIVVSSSSSKVNPLKTEAVVSSGLADRLRLGSLTEDGLSYKEKFIVRCYEVGINKTATVETIANLLQEVGCNHAQSVGYSTGGFSTTPTMRKLRLIWVTARMHIEIYKYPAWSDVVEIESWGQGEGKIGTRRDWILRDYATGQVIGRATSKWVMMNQDTRRLQKVDVDVRDEYLVHCPRELRLAFPEENNSSLKKISKLEDPSQYSKLGLVPRRADLDMNQHVNNVTYIGWVLESMPQEIIDTHELQTITLDYRRECQHDDVVDSLTSPEPSEDAEAVFNHNGTNGSANVSANDHGCRNFLHLLRLSGNGLEINRGRTEWRKKPTRMDYKDHDGDYKDHDIDYKDDDDKSEQ ID NO:137 Amino acid sequence of Gm FATA S111A, V193A mutant gene;D3998, pSZ5084. The algal transit peptide is underlined, the FLAG epitope tag isuppercase bold and the S111A, V193A residues are lower-case bold.MATASTFSAFNARCGDLRRSAGSGPRRPARPLPVRGRAIPPRIIVVSSSSSKVNPLKTEAVVSSGLADRLRLGSLTEDGLSYKEKFIVRCYEVGINKTATVETIANLLQEVGCNHAQSVGYSTGGFaTTPTMRKLRLIWVTARMHIEIYKYPAWSDVVEIESWGQGEGKIGTRRDWILRDYATGQVIGRATSKWVMMNQDTRRLQKVDaDVRDEYLVHCPRELRLAFPEENNSSLKKISKLEDPSQYSKLGLVPRRADLDMNQHVNNVTYIGWVLESMPQEIIDTHELQTITLDYRRECQHDDVVDSLTSPEPSEDAEAVFNHNGTNGSANVSANDHGCRNFLHLLRLSGNGLEINRGRTEWRKKPTRMDYKDHDGDYKDHDIDYKDDDDKSEQ ID NO:138 Amino acid sequence of Gm FATA S111V, V193A mutant gene;D3999, pSZ5085. The algal transit peptide is underlined, the FLAG epitope tag isuppercase bold and the S111V, V193A residues are lower-case bold.MATASTFSAFNARCGDLRRSAGSGPRRPARPLPVRGRAIPPRIIVVSSSSSKVNPLKTEAVVSSGLADRLRLGSLTEDGLSYKEKFIVRCYEVGINKTATVETIANLLQEVGCNHAQSVGYSTGGFvTTPTMRKLRLIWVTARMHIEIYKYPAWSDVVEIESWGQGEGKIGTRRDWILRDYATGQVIGRATSKWVMMNQDTRRLQKVDaDVRDEYLVHCPRELRLAFPEENNSSLKKISKLEDPSQYSKLGLVPRRADLDMNQHVNNVTYIGWVLESMPQEIIDTHELQTITLDYRRECQHDDVVDSLTSPEPSEDAEAVFNHNGTNGSANVSANDHGCRNFLHLLRLSGNGLEINRGRTEWRKKPTRMDYKDHDGDYKDHDIDYKDDDDKSEQ ID NO: 139 Amino acid sequence of Gm FATA G96A mutant gene; D4000,pSZ5086. The algal transit peptide is underlined, the FLAG epitope tag is uppercasebold and the G96A residue is lower-case bold.MATASTFSAFNARCGDLRRSAGSGPRRPARPLPVRGRAIPPRIIVVSSSSSKVNPLKTEAVVSSGLADRLRLGSLTEDGLSYKEKFIVRCYEVGINKTATVETIANLLQEVaCNHAQSVGYSTGGFSTTPTMRKLRLIWVTARMHIEIYKYPAWSDVVEIESWGQGEGKIGTRRDWILRDYATGQVIGRATSKWVMMNQDTRRLQKVDVDVRDEYLVHCPRELRLAFPEENNSSLKKISKLEDPSQYSKLGLVPRRADLDMNQHVNNVTYIGWVLESMPQEIIDTHELQTITLDYRRECQHDDVVDSLTSPEPSEDAEAVFNHNGTNGSANVSANDHGCRNFLHLLRLSGNGLEINRGRTEWRKKPTRMDYKDHDGDYKDHDIDYKDDDDKSEQ ID NO: 140 Amino acid sequence of Gm FATA G96T mutant gene; D4001,pSZ5087. The algal transit peptide is underlined, the FLAG epitope tag is uppercasebold and the G96T residue is lower-case bold.MATASTFSAFNARCGDLRRSAGSGPRRPARPLPVRGRAIPPRIIVVSSSSSKVNPLKTEAVVSSGLADRLRLGSLTEDGLSYKEKFIVRCYEVGINKTATVETIANLLQEVtCNHAQSVGYSTGGFSTTPTMRKLRLIWVTARMHIEIYKYPAWSDVVEIESWGQGEGKIGTRRDWILRDYATGQVIGRATSKWVMMNQDTRRLQKVDVDVRDEYLVHCPRELRLAFPEENNSSLKKISKLEDPSQYSKLGLVPRRADLDMNQHVNNVTYIGWVLESMPQEIIDTHELQTITLDYRRECQHDDVVDSLTSPEPSEDAEAVFNHNGTNGSANVSANDHGCRNFLHLLRLSGNGLEINRGRTEWRKKPTRMDYKDHDGDYKDHDIDYKDDDDKSEQ ID NO: 141 Amino acid sequence of Gm FATA G96V mutant gene; D4002,pSZ5088. The algal transit peptide is underlined, the FLAG epitope tag is uppercasebold and the G96V residue is lower-case bold.MATASTFSAFNARCGDLRRSAGSGPRRPARPLPVRGRAIPPRIIVVSSSSSKVNPLKTEAVVSSGLADRLRLGSLTEDGLSYKEKFIVRCYEVGINKTATVETIANLLQEVvCNHAQSVGYSTGGFSTTPTMRKLRLIWVTARMHIEIYKYPAWSDVVEIESWGQGEGKIGTRRDWILRDYATGQVIGRATSKWVMMNQDTRRLQKVDVDVRDEYLVHCPRELRLAFPEENNSSLKKISKLEDPSQYSKLGLVPRRADLDMNQHVNNVTYIGWVLESMPQEIIDTHELQTITLDYRRECQHDDVVDSLTSPEPSEDAEAVFNHNGTNGSANVSANDHGCRNFLHLLRLSGNGLEINRGRTEWRKKPTRMDYKDHDGDYKDHDIDYKDDDDKSEQ ID NO: 142 Amino acid sequence of Gm FATA G108A mutant gene;D4003, pSZ5089. The algal transit peptide is underlined, the FLAG epitope tag isuppercase bold and the G108A residue is lower-case bold.MATASTFSAFNARCGDLRRSAGSGPRRPARPLPVRGRAIPPRIIVVSSSSSKVNPLKTEAVVSSGLADRLRLGSLTEDGLSYKEKFIVRCYEVGINKTATVETIANLLQEVGCNHAQSVGYSTaGFSTTPTMRKLRLIWVTARMHIEIYKYPAWSDVVEIESWGQGEGKIGTRRDWILRDYATGQVIGRATSKWVMMNQDTRRLQKVDVDVRDEYLVHCPRELRLAFPEENNSSLKKISKLEDPSQYSKLGLVPRRADLDMNQHVNNVTYIGWVLESMPQEIIDTHELQTITLDYRRECQHDDVVDSLTSPEPSEDAEAVFNHNGTNGSANVSANDHGCRNFLHLLRLSGNGLEINRGRTEWRKKPTRMDYKDHDGDYKDHDIDYKDDDDKSEQ ID NO: 143 Amino acid sequence of Gm FATA L91F mutant gene; D4004,pSZ5090. The algal transit peptide is underlined, the FLAG epitope tag is uppercasebold and the L91F residue is lower-case bold.MATASTFSAFNARCGDLRRSAGSGPRRPARPLPVRGRAIPPRIIVVSSSSSKVNPLKTEAVVSSGLADRLRLGSLTEDGLSYKEKFIVRCYEVGINKTATVETIANfLQEVGCNHAQSVGYSTGGFSTTPTMRKLRLIWVTARMHIEIYKYPAWSDVVEIESWGQGEGKIGTRRDWILRDYATGQVIGRATSKWVMMNQDTRRLQKVDVDVRDEYLVHCPRELRLAFPEENNSSLKKISKLEDPSQYSKLGLVPRRADLDMNQHVNNVTYIGWVLESMPQEIIDTHELQTITLDYRRECQHDDVVDSLTSPEPSEDAEAVFNHNGTNGSANVSANDHGCRNFLHLLRLSGNGLEINRGRTEWRKKPTRMDYKDHDGDYKDHDIDYKDDDDKSEQ ID NO: 144 Amino acid sequence of Gm FATA L91K mutant gene; D4005,pSZ5091. The algal transit peptide is underlined, the FLAG epitope tag is uppercasebold and the L91K residue is lower-case boldMATASTFSAFNARCGDLRRSAGSGPRRPARPLPVRGRAIPPRIIVVSSSSSKVNPLKTEAVVSSGLADRLRLGSLTEDGLSYKEKFIVRCYEVGINKTATVETIANkLQEVGCNHAQSVGYSTGGFSTTPTMRKLRLIWVTARMHIEIYKYPAWSDVVEIESWGQGEGKIGTRRDWILRDYATGQVIGRATSKWVMMNQDTRRLQKVDVDVRDEYLVHCPRELRLAFPEENNSSLKKISKLEDPSQYSKLGLVPRRADLDMNQHVNNVTYIGWVLESMPQEIIDTHELQTITLDYRRECQHDDVVDSLTSPEPSEDAEAVFNHNGTNGSANVSANDHGCRNFLHLLRLSGNGLEINRGRTEWRKKPTRMDYKDHDGDYKDHDIDYKDDDDKSEQ ID NO:145 FIG. 10. Amino acid sequence of Gm FATA L915 mutantgene; D4006, pSZ5092. The algal transit peptide is underlined, the FLAG epitope tag isuppercase bold and the L91S residue is lower-case boldMATASTFSAFNARCGDLRRSAGSGPRRPARPLPVRGRAIPPRIIVVSSSSSKVNPLKTEAVVSSGLADRLRLGSLTEDGLSYKEKFIVRCYEVGINKTATVETIANsLQEVGCNHAQSVGYSTGGFSTTPTMRKLRLIWVTARMHIEIYKYPAWSDVVEIESWGQGEGKIGTRRDWILRDYATGQVIGRATSKWVMMNQDTRRLQKVDVDVRDEYLVHCPRELRLAFPEENNSSLKKISKLEDPSQYSKLGLVPRRADLDMNQHVNNVTYIGWVLESMPQEIIDTHELQTITLDYRRECQHDDVVDSLTSPEPSEDAEAVFNHNGTNGSANVSANDHGCRNFLHLLRLSGNGLEINRGRTEWRKKPTRMDYKDHDGDYKDHDIDYKDDDDKSEQ ID NO: 146 Amino acid sequence of Gm FATA G108V mutant gene;D4007, pSZ5093. The algal transit peptide is underlined, the FLAG epitope tag isuppercase bold and the G108V residue is lower-case bold.MATASTFSAFNARCGDLRRSAGSGPRRPARPLPVRGRAIPPRIIVVSSSSSKVNPLKTEAVVSSGLADRLRLGSLTEDGLSYKEKFIVRCYEVGINKTATVETIANLLQEVGCNHAQSVGYSTvGFSTTPTMRKLRLIWVTARMHIEIYKYPAWSDVVEIESWGQGEGKIGTRRDWILRDYATGQVIGRATSKWVMMNQDTRRLQKVDVDVRDEYLVHCPRELRLAFPEENNSSLKKISKLEDPSQYSKLGLVPRRADLDMNQHVNNVTYIGWVLESMPQEIIDTHELQTITLDYRRECQHDDVVDSLTSPEPSEDAEAVFNHNGTNGSANVSANDHGCRNFLHLLRLSGNGLEINRGRTEWRKKPTRMDYKDHDGDYKDHDIDYKDDDDKSEQ ID NO: 147 Amino acid sequence of Gm FATA T156F mutant gene;D4008, pSZ5094. The algal transit peptide is underlined, the FLAG epitope tag isuppercase bold and the T156F residue is lower-case bold.MATASTFSAFNARCGDLRRSAGSGPRRPARPLPVRGRAIPPRIIVVSSSSSKVNPLKTEAVVSSGLADRLRLGSLTEDGLSYKEKFIVRCYEVGINKTATVETIANLLQEVGCNHAQSVGYSTGGFSTTPTMRKLRLIWVTARMHIEIYKYPAWSDVVEIESWGQGEGKIGfRRDWILRDYATGQVIGRATSKWVMMNQDTRRLQKVDVDVRDEYLVHCPRELRLAFPEENNSSLKKISKLEDPSQYSKLGLVPRRADLDMNQHVNNVTYIGWVLESMPQEIIDTHELQTITLDYRRECQHDDVVDSLTSPEPSEDAEAVFNHNGTNGSANVSANDHGCRNFLHLLRLSGNGLEINRGRTEWRKKPTRMDYKDHDGDYKDHDIDYKDDDDKSEQ ID NO: 148 Amino acid sequence of Gm FATA T156A mutant gene;D4009, pSZ5095. The algal transit peptide is underlined, the FLAG epitope tag isuppercase bold and the T156A residue is lower-case bold.MATASTFSAFNARCGDLRRSAGSGPRRPARPLPVRGRAIPPRIIVVSSSSSKVNPLKTEAVVSSGLADRLRLGSLTEDGLSYKEKFIVRCYEVGINKTATVETIANLLQEVGCNHAQSVGYSTGGFSTTPTMRKLRLIWVTARMHIEIYKYPAWSDVVEIESWGQGEGKIGaRRDWILRDYATGQVIGRATSKWVMMNQDTRRLQKVDVDVRDEYLVHCPRELRLAFPEENNSSLKKISKLEDPSQYSKLGLVPRRADLDMNQHVNNVTYIGWVLESMPQEIIDTHELQTITLDYRRECQHDDVVDSLTSPEPSEDAEAVFNHNGTNGSANVSANDHGCRNFLHLLRLSGNGLEINRGRTEWRKKPTRMDYKDHDGDYKDHDIDYKDDDDKSEQ ID NO: 149 Amino acid sequence of Gm FATA T156K mutant gene; D4010,pSZ5096. The algal transit peptide is underlined, the FLAG epitope tag is uppercasebold and the T156K residue is lower-case bold.MATASTFSAFNARCGDLRRSAGSGPRRPARPLPVRGRAIPPRIIVVSSSSSKVNPLKTEAVVSSGLADRLRLGSLTEDGLSYKEKFIVRCYEVGINKTATVETIANLLQEVGCNHAQSVGYSTGGFSTTPTMRKLRLIWVTARMHIEIYKYPAWSDVVEIESWGQGEGKIGkRRDWILRDYATGQVIGRATSKWVMMNQDTRRLQKVDVDVRDEYLVHCPRELRLAFPEENNSSLKKISKLEDPSQYSKLGLVPRRADLDMNQHVNNVTYIGWVLESMPQEIIDTHELQTITLDYRRECQHDDVVDSLTSPEPSEDAEAVFNHNGTNGSANVSANDHGCRNFLHLLRLSGNGLEINRGRTEWRKKPTRMDYKDHDGDYKDHDIDYKDDDDKSEQ ID NO: 150 Amino acid sequence of Gm FATA T156V mutant gene;D4011, pSZ5097. The algal transit peptide is underlined, the FLAG epitope tag isuppercase bold and the T156V residue is lower-case bold.MATASTFSAFNARCGDLRRSAGSGPRRPARPLPVRGRAIPPRIIVVSSSSSKVNPLKTEAVVSSGLADRLRLGSLTEDGLSYKEKFIVRCYEVGINKTATVETIANLLQEVGCNHAQSVGYSTGGFSTTPTMRKLRLIWVTARMHIEIYKYPAWSDVVEIESWGQGEGKIGvRRDWILRDYATGQVIGRATSKWVMMNQDTRRLQKVDVDVRDEYLVHCPRELRLAFPEENNSSLKKISKLEDPSQYSKLGLVPRRADLDMNQHVNNVTYIGWVLESMPQEIIDTHELQTITLDYRRECQHDDVVDSLTSPEPSEDAEAVFNHNGTNGSANVSANDHGCRNFLHLLRLSGNGLEINRGRTEWRKKPTRMDYKDHDGDYKDHDIDYKDDDDKSEQ ID NO: 151 Nucleotide sequence of the GmFATA S111A, V193A mutant gene(D3998, pSZ5084). The promoter, 3′UTR, selection marker and targeting arms are the sameas pSZ5083.atggccaccgcatccactttctcggcgttcaatgcccgctgcggcgacctgcgtcgctcggcgggctccgggccccggcgcccagcgaggcccctccccgtgcgcgggcgcgccatccccccccgcatcatcgtggtgtcctcctcctcctccaaggtgaaccccctgaagaccgaggccgtggtgtcctccggcctggccgaccgcctgcgcctgggctccctgaccgaggacggcctgtcctacaaggagaagttcatcgtgcgctgctacgaggtgggcatcaacaagaccgccaccgtggagaccatcgccaacctgctgcaggaggtgggctgcaaccacgcccagtccgtgggctactccaccggcggcttcgccaccacccccaccatgcgcaagctgcgcctgatctgggtgaccgcccgcatgcacatcgagatctacaagtaccccgcctggtccgacgtggtggagatcgagtcctggggccagggcgagggcaagatcggcacccgccgcgactggatcctgcgcgactacgccaccggccaggtgatcggccgcgccacctccaagtgggtgatgatgaaccaggacacccgccgcctgcagaaggtggacgcggacgtgcgcgacgagtacctggtgcactgcccccgcgagctgcgcctggccttccccgaggagaacaactcctccctgaagaagatctccaagctggaggacccctcccagtactccaagctgggcctggtgccccgccgcgccgacctggacatgaaccagcacgtgaacaacgtgacctacatcggctgggtgctggagtccatgccccaggagatcatcgacacccacgagctgcagaccatcaccctggactaccgccgcgagtgccagcacgacgacgtggtggactccctgacctcccccgagccctccgaggacgccgaggccgtgttcaaccacaacggcaccaacggctccgccaacgtgtccgccaacgaccacggctgccgcaacttcctgcacctgctgcgcctgtccggcaacggcctggagatcaaccgcggccgcaccgagtggcgcaagaagcccacccgcatggactacaaggaccacgacggcgactacaaggaccacgacatcgactacaaggacgacgacgacaagtgaSEQ ID NO: 152 Nucleotide sequence of the GmFATA S111V, V193A mutant gene(D3999, pSZ5085). The promoter, 3′UTR, selection marker and targeting arms are thesame as pSZ5083.atggccaccgcatccactttctcggcgttcaatgcccgctgcggcgacctgcgtcgctcggcgggctccgggccccggcgcccagcgaggcccctccccgtgcgcgggcgcgccatccccccccgcatcatcgtggtgtcctcctcctcctccaaggtgaaccccctgaagaccgaggccgtggtgtcctccggcctggccgaccgcctgcgcctgggctccctgaccgaggacggcctgtcctacaaggagaagttcatcgtgcgctgctacgaggtgggcatcaacaagaccgccaccgtggagaccatcgccaacctgctgcaggaggtgggctgcaaccacgcccagtccgtgggctactccaccggcggcttcgtcaccacccccaccatgcgcaagctgcgcctgatctgggtgaccgcccgcatgcacatcgagatctacaagtaccccgcctggtccgacgtggtggagatcgagtcctggggccagggcgagggcaagatcggcacccgccgcgactggatcctgcgcgactacgccaccggccaggtgatcggccgcgccacctccaagtgggtgatgatgaaccaggacacccgccgcctgcagaaggtggacgcggacgtgcgcgacgagtacctggtgcactgcccccgcgagctgcgcctggccttccccgaggagaacaactcctccctgaagaagatctccaagctggaggacccctcccagtactccaagctgggcctggtgccccgccgcgccgacctggacatgaaccagcacgtgaacaacgtgacctacatcggctgggtgctggagtccatgccccaggagatcatcgacacccacgagctgcagaccatcaccctggactaccgccgcgagtgccagcacgacgacgtggtggactccctgacctcccccgagccctccgaggacgccgaggccgtgttcaaccacaacggcaccaacggctccgccaacgtgtccgccaacgaccacggctgccgcaacttcctgcacctgctgcgcctgtccggcaacggcctggagatcaaccgcggccgcaccgagtggcgcaagaagcccacccgcatggactacaaggaccacgacggcgactacaaggaccacgacatcgactacaaggacgacgacgacaagtgaSEQ ID NO: 153 Nucleotide sequence of the GmFATA G96A mutant gene (D4000,pSZ5086). The promoter, 3′UTR, selection marker and targeting arms are the same aspSZ5083 atggccaccgcatccactttctcggcgttcaatgcccgctgcggcgacctgcgtcgctcggcgggctccgggccccggcgcccagcgaggcccctccccgtgcgcgggcgcgccatccccccccgcatcatcgtggtgtcctcctcctcctccaaggtgaaccccctgaagaccgaggccgtggtgtcctccggcctggccgaccgcctgcgcctgggctccctgaccgaggacggcctgtcctacaaggagaagttcatcgtgcgctgctacgaggtgggcatcaacaagaccgccaccgtggagaccatcgccaacctgctgcaggaggtggcgtgcaaccacgcccagtccgtgggctactccaccggcggcttctccaccacccccaccatgcgcaagctgcgcctgatctgggtgaccgcccgcatgcacatcgagatctacaagtaccccgcctggtccgacgtggtggagatcgagtcctggggccagggcgagggcaagatcggcacccgccgcgactggatcctgcgcgactacgccaccggccaggtgatcggccgcgccacctccaagtgggtgatgatgaaccaggacacccgccgcctgcagaaggtggacgtggacgtgcgcgacgagtacctggtgcactgcccccgcgagctgcgcctggccttccccgaggagaacaactcctccctgaagaagatctccaagctggaggacccctcccagtactccaagctgggcctggtgccccgccgcgccgacctggacatgaaccagcacgtgaacaacgtgacctacatcggctgggtgctggagtccatgccccaggagatcatcgacacccacgagctgcagaccatcaccctggactaccgccgcgagtgccagcacgacgacgtggtggactccctgacctcccccgagccctccgaggacgccgaggccgtgttcaaccacaacggcaccaacggctccgccaacgtgtccgccaacgaccacggctgccgcaacttcctgcacctgctgcgcctgtccggcaacggcctggagatcaaccgcggccgcaccgagtggcgcaagaagcccacccgcatggactacaaggaccacgacggcgactacaaggaccacgacatcgactacaaggacgacgacgacaagtgaSEQ ID NO: 154 Nucleotide sequence of the GmFATA G96T mutant gene (D4001,pSZ5087). The promoter, 3′UTR, selection marker and targeting arms are the same aspSZ5083 atggccaccgcatccactttctcggcgttcaatgcccgctgcggcgacctgcgtcgctcggcgggctccgggccccggcgcccagcgaggcccctccccgtgcgcgggcgcgccatccccccccgcatcatcgtggtgtcctcctcctcctccaaggtgaaccccctgaagaccgaggccgtggtgtcctccggcctggccgaccgcctgcgcctgggctccctgaccgaggacggcctgtcctacaaggagaagttcatcgtgcgctgctacgaggtgggcatcaacaagaccgccaccgtggagaccatcgccaacctgctgcaggaggtgacgtgcaaccacgcccagtccgtgggctactccaccggcggcttctccaccacccccaccatgcgcaagctgcgcctgatctgggtgaccgcccgcatgcacatcgagatctacaagtaccccgcctggtccgacgtggtggagatcgagtcctggggccagggcgagggcaagatcggcacccgccgcgactggatcctgcgcgactacgccaccggccaggtgatcggccgcgccacctccaagtgggtgatgatgaaccaggacacccgccgcctgcagaaggtggacgtggacgtgcgcgacgagtacctggtgcactgcccccgcgagctgcgcctggccttccccgaggagaacaactcctccctgaagaagatctccaagctggaggacccctcccagtactccaagctgggcctggtgccccgccgcgccgacctggacatgaaccagcacgtgaacaacgtgacctacatcggctgggtgctggagtccatgccccaggagatcatcgacacccacgagctgcagaccatcaccctggactaccgccgcgagtgccagcacgacgacgtggtggactccctgacctcccccgagccctccgaggacgccgaggccgtgttcaaccacaacggcaccaacggctccgccaacgtgtccgccaacgaccacggctgccgcaacttcctgcacctgctgcgcctgtccggcaacggcctggagatcaaccgcggccgcaccgagtggcgcaagaagcccacccgcatggactacaaggaccacgacggcgactacaaggaccacgacatcgactacaaggacgacgacgacaagtgaSEQ ID NO: 155 Nucleotide sequence of the GmFATA G96V mutant gene (D4002,pSZ5088). The promoter, 3′UTR, selection marker and targeting arms are the same aspSZ5083. atggccaccgcatccactttctcggcgttcaatgcccgctgcggcgacctgcgtcgctcggcgggctccgggccccggcgcccagcgaggcccctccccgtgcgcgggcgcgccatccccccccgcatcatcgtggtgtcctcctcctcctccaaggtgaaccccctgaagaccgaggccgtggtgtcctccggcctggccgaccgcctgcgcctgggctccctgaccgaggacggcctgtcctacaaggagaagttcatcgtgcgctgctacgaggtgggcatcaacaagaccgccaccgtggagaccatcgccaacctgctgcaggaggtggtgtgcaaccacgcccagtccgtgggctactccaccggcggcttctccaccacccccaccatgcgcaagctgcgcctgatctgggtgaccgcccgcatgcacatcgagatctacaagtaccccgcctggtccgacgtggtggagatcgagtcctggggccagggcgagggcaagatcggcacccgccgcgactggatcctgcgcgactacgccaccggccaggtgatcggccgcgccacctccaagtgggtgatgatgaaccaggacacccgccgcctgcagaaggtggacgtggacgtgcgcgacgagtacctggtgcactgcccccgcgagctgcgcctggccttccccgaggagaacaactcctccctgaagaagatctccaagctggaggacccctcccagtactccaagctgggcctggtgccccgccgcgccgacctggacatgaaccagcacgtgaacaacgtgacctacatcggctgggtgctggagtccatgccccaggagatcatcgacacccacgagctgcagaccatcaccctggactaccgccgcgagtgccagcacgacgacgtggtggactccctgacctcccccgagccctccgaggacgccgaggccgtgttcaaccacaacggcaccaacggctccgccaacgtgtccgccaacgaccacggctgccgcaacttcctgcacctgctgcgcctgtccggcaacggcctggagatcaaccgcggccgcaccgagtggcgcaagaagcccacccgcatggactacaaggaccacgacggcgactacaaggaccacgacatcgactacaaggacgacgacgacaagtgaSEQ ID NO: 156 Nucleotide sequence of the GmFATA G108A mutant gene(D4003, pSZ5089). The promoter, 3′UTR, selection marker and targeting arms are thesame as pSZ50836.atggccaccgcatccactttctcggcgttcaatgcccgctgcggcgacctgcgtcgctcggcgggctccgggccccggcgcccagcgaggcccctccccgtgcgcgggcgcgccatccccccccgcatcatcgtggtgtcctcctcctcctccaaggtgaaccccctgaagaccgaggccgtggtgtcctccggcctggccgaccgcctgcgcctgggctccctgaccgaggacggcctgtcctacaaggagaagttcatcgtgcgctgctacgaggtgggcatcaacaagaccgccaccgtggagaccatcgccaacctgctgcaggaggtgggctgcaaccacgcccagtccgtgggctactccaccgccggcttctccaccacccccaccatgcgcaagctgcgcctgatctgggtgaccgcccgcatgcacatcgagatctacaagtaccccgcctggtccgacgtggtggagatcgagtcctggggccagggcgagggcaagatcggcacccgccgcgactggatcctgcgcgactacgccaccggccaggtgatcggccgcgccacctccaagtgggtgatgatgaaccaggacacccgccgcctgcagaaggtggacgtggacgtgcgcgacgagtacctggtgcactgcccccgcgagctgcgcctggccttccccgaggagaacaactcctccctgaagaagatctccaagctggaggacccctcccagtactccaagctgggcctggtgccccgccgcgccgacctggacatgaaccagcacgtgaacaacgtgacctacatcggctgggtgctggagtccatgccccaggagatcatcgacacccacgagctgcagaccatcaccctggactaccgccgcgagtgccagcacgacgacgtggtggactccctgacctcccccgagccctccgaggacgccgaggccgtgttcaaccacaacggcaccaacggctccgccaacgtgtccgccaacgaccacggctgccgcaacttcctgcacctgctgcgcctgtccggcaacggcctggagatcaaccgcggccgcaccgagtggcgcaagaagcccacccgcatggactacaaggaccacgacggcgactacaaggaccacgacatcgactacaaggacgacgacgacaagtgaSEQ ID NO: 157 Nucleotide sequence of the GmFATA L91F mutant gene (D4004,pSZ5090). The promoter, 3′UTR, selection marker and targeting arms are the same aspSZ5083 atggccaccgcatccactttctcggcgttcaatgcccgctgcggcgacctgcgtcgctcggcgggctccgggccccggcgcccagcgaggcccctccccgtgcgcgggcgcgccatccccccccgcatcatcgtggtgtcctcctcctcctccaaggtgaaccccctgaagaccgaggccgtggtgtcctccggcctggccgaccgcctgcgcctgggctccctgaccgaggacggcctgtcctacaaggagaagttcatcgtgcgctgctacgaggtgggcatcaacaagaccgccaccgtggagaccatcgccaacttcctgcaggaggtgggctgcaaccacgcccagtccgtgggctactccaccggcggcttctccaccacccccaccatgcgcaagctgcgcctgatctgggtgaccgcccgcatgcacatcgagatctacaagtaccccgcctggtccgacgtggtggagatcgagtcctggggccagggcgagggcaagatcggcacccgccgcgactggatcctgcgcgactacgccaccggccaggtgatcggccgcgccacctccaagtgggtgatgatgaaccaggacacccgccgcctgcagaaggtggacgtggacgtgcgcgacgagtacctggtgcactgcccccgcgagctgcgcctggccttccccgaggagaacaactcctccctgaagaagatctccaagctggaggacccctcccagtactccaagctgggcctggtgccccgccgcgccgacctggacatgaaccagcacgtgaacaacgtgacctacatcggctgggtgctggagtccatgccccaggagatcatcgacacccacgagctgcagaccatcaccctggactaccgccgcgagtgccagcacgacgacgtggtggactccctgacctcccccgagccctccgaggacgccgaggccgtgttcaaccacaacggcaccaacggctccgccaacgtgtccgccaacgaccacggctgccgcaacttcctgcacctgctgcgcctgtccggcaacggcctggagatcaaccgcggccgcaccgagtggcgcaagaagcccacccgcatggactacaaggaccacgacggcgactacaaggaccacgacatcgactacaaggacgacgacgacaagtgaSEQ ID NO: 158 Nucleotide sequence of the GmFATA L91K mutant gene (D4005,pSZ5091). The promoter, 3′UTR, selection marker and targeting arms are the same aspSZ5083. atggccaccgcatccactttctcggcgttcaatgcccgctgcggcgacctgcgtcgctcggcgggctccgggccccggcgcccagcgaggcccctccccgtgcgcgggcgcgccatccccccccgcatcatcgtggtgtcctcctcctcctccaaggtgaaccccctgaagaccgaggccgtggtgtcctccggcctggccgaccgcctgcgcctgggctccctgaccgaggacggcctgtcctacaaggagaagttcatcgtgcgctgctacgaggtgggcatcaacaagaccgccaccgtggagaccatcgccaacaagctgcaggaggtgggctgcaaccacgcccagtccgtgggctactccaccggcggcttctccaccacccccaccatgcgcaagctgcgcctgatctgggtgaccgcccgcatgcacatcgagatctacaagtaccccgcctggtccgacgtggtggagatcgagtcctggggccagggcgagggcaagatcggcacccgccgcgactggatcctgcgcgactacgccaccggccaggtgatcggccgcgccacctccaagtgggtgatgatgaaccaggacacccgccgcctgcagaaggtggacgtggacgtgcgcgacgagtacctggtgcactgcccccgcgagctgcgcctggccttccccgaggagaacaactcctccctgaagaagatctccaagctggaggacccctcccagtactccaagctgggcctggtgccccgccgcgccgacctggacatgaaccagcacgtgaacaacgtgacctacatcggctgggtgctggagtccatgccccaggagatcatcgacacccacgagctgcagaccatcaccctggactaccgccgcgagtgccagcacgacgacgtggtggactccctgacctcccccgagccctccgaggacgccgaggccgtgttcaaccacaacggcaccaacggctccgccaacgtgtccgccaacgaccacggctgccgcaacttcctgcacctgctgcgcctgtccggcaacggcctggagatcaaccgcggccgcaccgagtggcgcaagaagcccacccgcatggactacaaggaccacgacggcgactacaaggaccacgacatcgactacaaggacgacgacgacaagtgaSEQ ID NO: 159 Nucleotide sequence of the GmFATA L91S mutant gene (D4006,pSZ5092). The promoter, 3′UTR, selection marker and targeting arms are the same aspSZ5083. atggccaccgcatccactttctcggcgttcaatgcccgctgcggcgacctgcgtcgctcggcgggctccgggccccggcgcccagcgaggcccctccccgtgcgcgggcgcgccatccccccccgcatcatcgtggtgtcctcctcctcctccaaggtgaaccccctgaagaccgaggccgtggtgtcctccggcctggccgaccgcctgcgcctgggctccctgaccgaggacggcctgtcctacaaggagaagttcatcgtgcgctgctacgaggtgggcatcaacaagaccgccaccgtggagaccatcgccaactcgctgcaggaggtgggctgcaaccacgcccagtccgtgggctactccaccggcggcttctccaccacccccaccatgcgcaagctgcgcctgatctgggtgaccgcccgcatgcacatcgagatctacaagtaccccgcctggtccgacgtggtggagatcgagtcctggggccagggcgagggcaagatcggcacccgccgcgactggatcctgcgcgactacgccaccggccaggtgatcggccgcgccacctccaagtgggtgatgatgaaccaggacacccgccgcctgcagaaggtggacgtggacgtgcgcgacgagtacctggtgcactgcccccgcgagctgcgcctggccttccccgaggagaacaactcctccctgaagaagatctccaagctggaggacccctcccagtactccaagctgggcctggtgccccgccgcgccgacctggacatgaaccagcacgtgaacaacgtgacctacatcggctgggtgctggagtccatgccccaggagatcatcgacacccacgagctgcagaccatcaccctggactaccgccgcgagtgccagcacgacgacgtggtggactccctgacctcccccgagccctccgaggacgccgaggccgtgttcaaccacaacggcaccaacggctccgccaacgtgtccgccaacgaccacggctgccgcaacttcctgcacctgctgcgcctgtccggcaacggcctggagatcaaccgcggccgcaccgagtggcgcaagaagcccacccgcatggactacaaggaccacgacggcgactacaaggaccacgacatcgactacaaggacgacgacgacaagtgaSEQ ID NO: 160 Nucleotide sequence of the GmFATA G108V mutant gene(D4007, pSZ5093). The promoter, 3′UTR, selection marker and targeting arms are thesame as pSZ5083.atggccaccgcatccactttctcggcgttcaatgcccgctgcggcgacctgcgtcgctcggcgggctccgggccccggcgcccagcgaggcccctccccgtgcgcgggcgcgccatccccccccgcatcatcgtggtgtcctcctcctcctccaaggtgaaccccctgaagaccgaggccgtggtgtcctccggcctggccgaccgcctgcgcctgggctccctgaccgaggacggcctgtcctacaaggagaagttcatcgtgcgctgctacgaggtgggcatcaacaagaccgccaccgtggagaccatcgccaacctgctgcaggaggtgggctgcaaccacgcccagtccgtgggctactccaccgtcggcttctccaccacccccaccatgcgcaagctgcgcctgatctgggtgaccgcccgcatgcacatcgagatctacaagtaccccgcctggtccgacgtggtggagatcgagtcctggggccagggcgagggcaagatcggcacccgccgcgactggatcctgcgcgactacgccaccggccaggtgatcggccgcgccacctccaagtgggtgatgatgaaccaggacacccgccgcctgcagaaggtggacgtggacgtgcgcgacgagtacctggtgcactgcccccgcgagctgcgcctggccttccccgaggagaacaactcctccctgaagaagatctccaagctggaggacccctcccagtactccaagctgggcctggtgccccgccgcgccgacctggacatgaaccagcacgtgaacaacgtgacctacatcggctgggtgctggagtccatgccccaggagatcatcgacacccacgagctgcagaccatcaccctggactaccgccgcgagtgccagcacgacgacgtggtggactccctgacctcccccgagccctccgaggacgccgaggccgtgttcaaccacaacggcaccaacggctccgccaacgtgtccgccaacgaccacggctgccgcaacttcctgcacctgctgcgcctgtccggcaacggcctggagatcaaccgcggccgcaccgagtggcgcaagaagcccacccgcatggactacaaggaccacgacggcgactacaaggaccacgacatcgactacaaggacgacgacgacaagtgaSEQ ID NO: 161 Nucleotide sequence of the GmFATA T156F mutant gene (D4008,pSZ5094). The promoter, 3′UTR, selection marker and targeting arms are the same aspSZ5083. atggccaccgcatccactttctcggcgttcaatgcccgctgcggcgacctgcgtcgctcggcgggctccgggccccggcgcccagcgaggcccctccccgtgcgcgggcgcgccatccccccccgcatcatcgtggtgtcctcctcctcctccaaggtgaaccccctgaagaccgaggccgtggtgtcctccggcctggccgaccgcctgcgcctgggctccctgaccgaggacggcctgtcctacaaggagaagttcatcgtgcgctgctacgaggtgggcatcaacaagaccgccaccgtggagaccatcgccaacctgctgcaggaggtgggctgcaaccacgcccagtccgtgggctactccaccggcggcttctccaccacccccaccatgcgcaagctgcgcctgatctgggtgaccgcccgcatgcacatcgagatctacaagtaccccgcctggtccgacgtggtggagatcgagtcctggggccagggcgagggcaagatcggcttccgccgcgactggatcctgcgcgactacgccaccggccaggtgatcggccgcgccacctccaagtgggtgatgatgaaccaggacacccgccgcctgcagaaggtggacgtggacgtgcgcgacgagtacctggtgcactgcccccgcgagctgcgcctggccttccccgaggagaacaactcctccctgaagaagatctccaagctggaggacccctcccagtactccaagctgggcctggtgccccgccgcgccgacctggacatgaaccagcacgtgaacaacgtgacctacatcggctgggtgctggagtccatgccccaggagatcatcgacacccacgagctgcagaccatcaccctggactaccgccgcgagtgccagcacgacgacgtggtggactccctgacctcccccgagccctccgaggacgccgaggccgtgttcaaccacaacggcaccaacggctccgccaacgtgtccgccaacgaccacggctgccgcaacttcctgcacctgctgcgcctgtccggcaacggcctggagatcaaccgcggccgcaccgagtggcgcaagaagcccacccgcatggactacaaggaccacgacggcgactacaaggaccacgacatcgactacaaggacgacgacgacaagtgaSEQ ID NO: 162 Nucleotide sequence of the GmFATA T156A mutant gene (D4009,pSZ5095). The promoter, 3′UTR, selection marker and targeting arms are the same aspSZ5083 atggccaccgcatccactttctcggcgttcaatgcccgctgcggcgacctgcgtcgctcggcgggctccgggccccggcgcccagcgaggcccctccccgtgcgcgggcgcgccatccccccccgcatcatcgtggtgtcctcctcctcctccaaggtgaaccccctgaagaccgaggccgtggtgtcctccggcctggccgaccgcctgcgcctgggctccctgaccgaggacggcctgtcctacaaggagaagttcatcgtgcgctgctacgaggtgggcatcaacaagaccgccaccgtggagaccatcgccaacctgctgcaggaggtgggctgcaaccacgcccagtccgtgggctactccaccggcggcttctccaccacccccaccatgcgcaagctgcgcctgatctgggtgaccgcccgcatgcacatcgagatctacaagtaccccgcctggtccgacgtggtggagatcgagtcctggggccagggcgagggcaagatcggcgcgcgccgcgactggatcctgcgcgactacgccaccggccaggtgatcggccgcgccacctccaagtgggtgatgatgaaccaggacacccgccgcctgcagaaggtggacgtggacgtgcgcgacgagtacctggtgcactgcccccgcgagctgcgcctggccttccccgaggagaacaactcctccctgaagaagatctccaagctggaggacccctcccagtactccaagctgggcctggtgccccgccgcgccgacctggacatgaaccagcacgtgaacaacgtgacctacatcggctgggtgctggagtccatgccccaggagatcatcgacacccacgagctgcagaccatcaccctggactaccgccgcgagtgccagcacgacgacgtggtggactccctgacctcccccgagccctccgaggacgccgaggccgtgttcaaccacaacggcaccaacggctccgccaacgtgtccgccaacgaccacggctgccgcaacttcctgcacctgctgcgcctgtccggcaacggcctggagatcaaccgcggccgcaccgagtggcgcaagaagcccacccgcatggactacaaggaccacgacggcgactacaaggaccacgacatcgactacaaggacgacgacgacaagtgaSEQ ID NO: 163 Nucleotide sequence of the GmFATA T156K mutant gene (D4010,pSZ5096). The promoter, 3′UTR, selection marker and targeting arms are the same aspSZ5083. atggccaccgcatccactttctcggcgttcaatgcccgctgcggcgacctgcgtcgctcggcgggctccgggccccggcgcccagcgaggcccctccccgtgcgcgggcgcgccatccccccccgcatcatcgtggtgtcctcctcctcctccaaggtgaaccccctgaagaccgaggccgtggtgtcctccggcctggccgaccgcctgcgcctgggctccctgaccgaggacggcctgtcctacaaggagaagttcatcgtgcgctgctacgaggtgggcatcaacaagaccgccaccgtggagaccatcgccaacctgctgcaggaggtgggctgcaaccacgcccagtccgtgggctactccaccggcggcttctccaccacccccaccatgcgcaagctgcgcctgatctgggtgaccgcccgcatgcacatcgagatctacaagtaccccgcctggtccgacgtggtggagatcgagtcctggggccagggcgagggcaagatcggcaagcgccgcgactggatcctgcgcgactacgccaccggccaggtgatcggccgcgccacctccaagtgggtgatgatgaaccaggacacccgccgcctgcagaaggtggacgtggacgtgcgcgacgagtacctggtgcactgcccccgcgagctgcgcctggccttccccgaggagaacaactcctccctgaagaagatctccaagctggaggacccctcccagtactccaagctgggcctggtgccccgccgcgccgacctggacatgaaccagcacgtgaacaacgtgacctacatcggctgggtgctggagtccatgccccaggagatcatcgacacccacgagctgcagaccatcaccctggactaccgccgcgagtgccagcacgacgacgtggtggactccctgacctcccccgagccctccgaggacgccgaggccgtgttcaaccacaacggcaccaacggctccgccaacgtgtccgccaacgaccacggctgccgcaacttcctgcacctgctgcgcctgtccggcaacggcctggagatcaaccgcggccgcaccgagtggcgcaagaagcccacccgcatggactacaaggaccacgacggcgactacaaggaccacgacatcgactacaaggacgacgacgacaagtgaSEQ ID NO: 164 Nucleotide sequence of the GmFATA T156V mutant gene (D4011,pSZ5097). The promoter, 3′UTR, selection marker and targeting arms are the same aspSZ5083 atggccaccgcatccactttctcggcgttcaatgcccgctgcggcgacctgcgtcgctcggcgggctccgggccccggcgcccagcgaggcccctccccgtgcgcgggcgcgccatccccccccgcatcatcgtggtgtcctcctcctcctccaaggtgaaccccctgaagaccgaggccgtggtgtcctccggcctggccgaccgcctgcgcctgggctccctgaccgaggacggcctgtcctacaaggagaagttcatcgtgcgctgctacgaggtgggcatcaacaagaccgccaccgtggagaccatcgccaacctgctgcaggaggtgggctgcaaccacgcccagtccgtgggctactccaccggcggcttctccaccacccccaccatgcgcaagctgcgcctgatctgggtgaccgcccgcatgcacatcgagatctacaagtaccccgcctggtccgacgtggtggagatcgagtcctggggccagggcgagggcaagatcggcgtgcgccgcgactggatcctgcgcgactacgccaccggccaggtgatcggccgcgccacctccaagtgggtgatgatgaaccaggacacccgccgcctgcagaaggtggacgtggacgtgcgcgacgagtacctggtgcactgcccccgcgagctgcgcctggccttccccgaggagaacaactcctccctgaagaagatctccaagctggaggacccctcccagtactccaagctgggcctggtgccccgccgcgccgacctggacatgaaccagcacgtgaacaacgtgacctacatcggctgggtgctggagtccatgccccaggagatcatcgacacccacgagctgcagaccatcaccctggactaccgccgcgagtgccagcacgacgacgtggtggactccctgacctcccccgagccctccgaggacgccgaggccgtgttcaaccacaacggcaccaacggctccgccaacgtgtccgccaacgaccacggctgccgcaacttcctgcacctgctgcgcctgtccggcaacggcctggagatcaaccgcggccgcaccgagtggcgcaagaagcccacccgcatggactacaaggaccacgacggcgactacaaggaccacgacatcgactacaaggacgacgacgacaagtga

SEQUENCES SEQ ID NO: 1 gcgaggggtc tgcctgggcc agccgctccc tctgaacacgggacgcgtgg tccaattcgg 60 gcttcgggac cctttggcgg tttgaacgcc tgggagagggcgcccgcgag cctggggacc 120 ccggcaacgg cttccccaga gcctgccttg caatctcgcgcgtcctctcc ctcagcacgt 180 ggcggttcca cgtgtggtcg ggcgtcccgg actagctcacgtcgtgacct agcttaatga 240 acccagccgg gcctgcagca ccaccttaga ggttttgattatttgattag accaatctat 300 tcacc 305 SEQ ID NO: 2 ggcgaataga ttggtataatgaaataatca aaacctctta ggcggtgcta caggcccggc 60 tgggttcatt aagctaggtcacgacgcgag ctagtccggg aagcccgacc acacgtggaa 120 ccgccacgtg ctgagggagaggacgcgcga gattgcaagg caggctctgg ggaagccgtt 180 gccggggtcc ccaggctcgcgggcgcccca tccctggcgt tcaaaccgcc aaagggtccc 240 gaagcccgaa ttggaccacgcgtcccgtgt ttagagggag cggctggccc aggcagaccc 300 ctcgc 305 SEQ ID NO: 3ggtgaataga ttggtctaat caaataatca aaacctctaa ggtggtgctg caggcccggc 60tgggttcatt aagctaggtc acgacgtgag ctagtccggg acgcccgacc acacgtggaa 120ccgccacgtg ctgagggaga ggacgcgcga gattgcaagg caggctctgg ggaagccgtt 180gccggggtcc ccaggctcgc gggcgccctc tcccaggcg tcaaaccgcc aaagggtccc 240gaagcccgaa ttggaccacg cgtcccgtgt tcagagggag cggctggccc aggcagaccc 300ctcgc 305 SEQ ID NO: 4 gtgatgggtt ctttagacga tccagcccag gatcatgtgttgcccacatg gagcctatcc 60 acgctggcct agaaggcaag cacatttcaa ggtgaacccacgtccatgga gcgatggcgc 120 caatatctcg cctctagacc aagcggttct caccccaactgcgtcatttg tatgtatggc 180 tgcaaagttg tcggtacgat agaggccgcc aacctggcggcgagggcgag gagctggttg 240 ccgatctgtg cccaagcatg tgtcggagct cggctgtctcggcagcgagc tcctgtgcaa 300 ggggcttgca tcgagaatgt caggcgatag acactgcacgttggggacac ggaggtgccc 360 ctgtggcgtg tcctggatgc cctcgggtcc gtcgcgagaagctctggcga ccagcacccg 420 gccacaaccg cagcaggcgt tcacccacaa gaatcttccagatcgtgatg cgcatgtatc 480 gtgacacgat tggcgaggtc cgcaggacgc acacggactcgtccactcat cagaactggt 540 cagggcaccc atctgcgtcc cttttcagga accacccaccgctgccaggc accttcgcca 600 gcggcggact ccacacagag aatgccttgc tgtgagagaccatggccggc aagtgctgtc 660 ggatctgccc gcatacggtc agtccccagc acaaggaagccaagagtaca ggctgttggt 720 gtcgatggag gagtggccgt tcccacaagt agtgagcggcagctgctcaa cggcttcccc 780 ctgttcatct tggcaaagcc agtgacttcc tacaagtatgtgatgcagat cggcactgca 840 atctgtcggc atgcgtacag aacatcggct cgccagggcagcgttgctcg ctctggatga 900 gctgcttggg aggaatcatc ggcacacgcc cgtgccgtgcccgcgccccg cgcccgtcgg 960 gaaaggcccc cggttaggac actgccgcgt cagccagtcgtgggatcgat cggacgtggc 1020 gaatcctcgc ccggacaccc tcatcacacc ccacatttccctgcaagcaa tcttgccgac 1080 aaaatagtca agatccattg ggtttaggga acacgtgcgagactgggcag ctgtatctgt 1140 ccttgccccg cgtcaaattc ctgggcgtga cgcagtcacaggagaatcta ttagaccctg 1200 gacttgcagc tcagtcatgg gcgtgagtgg ctaaagcacctaggtcaggc gagtaccgcc 1260 ccttccccag gattcactct tctgcgattg acgttgagcctgcatcgggc tgcttcgtca 1320 cc 1322 SEQ ID NO: 5 tcggagctaa agcagagactggacaagact tgcgttcgca tactggtgac acagaatagc 60 tcccatctat tcatacgcctttgggaaaag gaacgagcct tgtggcctct gcattgctgc 120 ctgctttgag gccgaggacggtgcgggacg ctcagatcca tcagcgatcg  ccccaccctc 180 agagcacctc cgatccaaggcaatactatc aggcaaagtt tccaaattca aacattccaa 240 aatcacgcca gggactggatcacacacgca gatcagcgcc gttttgctct ttgcctacgg 300 gcgactgtgc cacttgtcgacccctggtga cgggagggac cacgcctgcg gttggcatcc 360 acttcgacgg acccagggacggtttctcat gccaaacctg agatttgagc acccagatga 420 gcacattatg cgttttaggatgcctgagca gcgggcgtgc aggaatctgg tctcgccaga 480 ttcaccgaag atgcgcccatcggagcgagg cgagggcttt gtgaccacgc aaggcagtgt 540 gaggcaaaca catagggacacctgcgtctt tcaatgcaca gacatctatg gtgcccatgt 600 atataaaatg ggctacttctgagtcaaacc aacgcaaact gcgctatggc aaggccggcc 660 aaggttggaa tcccggtctgtctggatttg agtttgtggg ggctatcacg tgacaatccc 720 tgggattggg cggcagcagcgcacggcctg ggtggcaatg gcgcactaat actgctgaaa 780 gcacggctct gcatccctttctcttgacct gcgattggtc cttttcgcaa gcgtgatcat 840 c 841 SEQ ID NO: 6tcggagctaa agcagaaact gaacaagact tgcgttcgca tacttgtgac actgaatagg 60ttcaatctat tcatacgcct ttgggaaact gaacgagcct tgtggcctct gcattgctgc 120ctgctttgag gccgaggacg gcgcggaacg cacagatcca tcagcgatcg ccccaccctc 180agagtacatc cgatccaagg caatactatc aggcaaagtt tccaaattca aacattccaa 240aattacgtca gggactggat cacacacgca gatcagcgcc gttttgctct ttgcctacgg 300gcgactgtgc cacttgtcga cgcctggtga cgggagggac cacgcctgcg gttggcatcc 360acttcgacgg acccagggac ggtctcacat gccaaacctg agatttgagc accaagatga 420gcacattatg cgtttttgga tgcctgagca gcgggcgtgc aggaatctgg tctcgccaga 480ttcaccgaag atgcggccat cggagcgagg cgagggctgt gtggccacgc caggcagtgt 540gaggcaaaca cacagggaca tctgcttctt tcgatgcaca gacatctatg ttgcccgtgc 600atataaaatg ggctacttct gaatcaaacc aacgcaaact tcgctatggc aaggccggcc 660aaggttggaa tcccggtctg tctggatttg agtttgtggg ggctatcacg tgacaatccc 720tgggattggg cggcagcagc gcacggcctg gatggcaatg gcgcactaat actgctgaaa 780gcacggctct gcatcccttt ctcttgacct gcgattggtc cttttcgcaa gcgtgatcat 840 c841 SEQ ID NO: 7 caccgatcac tccgtcgccg cccaagagaa atcaacctcg atggagggcgaggtggatca 60 gaggtattgg ttatcgttcg ttcttagtct caatcaatcg tacaccttgcagttgcccga 120 gtttctccac acatacagca cctcccgctc ccagcccatt cgagcgacccaatccgggcg 180 atcccagcga tcgtcgtcgc ttcagtgctg accggtggaa agcaggagatctcgggcgag 240 caggaccaca tccagcccag gatcttcgac tggctcagag ctgaccctcacgcggcacag 300 caaaagtagc acgcacgcgt tatgcaaact ggttacaacc tgtccaacagtgttgcgacg 360 ttgactggct acattgtctg tctgtcgcga gtgcgcctgg gcccttacggtgggacactg 420 gaactccgcc ccgagtcgaa cacctagggc gacgcccgca gcttggcatgacagctctcc 480 ttgtgttcta aataccttgc gcgtgtggga  ga 512 SEQ ID NO: 8atccaccgat cactccgtcg ccgcccaaga gaattcaacc tcgatggagg gcaaggtgga 60tcagaggtat tggttatcgt tcgctattag tctcaatcaa tcgtgcacct tgcagttgct 120cgagtttctc cacacataca gcacctcccg ctcccagccc attcgagcga cccaatccgg 180gcgatcccag cgatcgtcgt cgcttcagtg ctgaccggtg gaaagcagga gatctcgggc 240gagcaggacc acatccagca caggatcttc gactggctca gagctgaccc tcacgcggca 300cagcaaaagt agcccgcacg cgttatgcaa acaggttaca acctgtccaa cactgttgcg 360acgttgactg gctacattgt ctgtctgtcg cgagtacgcc tggaccctta cggtgggaca 420ctggaactcc gccccgagtc gaacacctag ggcgacgccc gcagcttggc atgacagctc 480tccttgtatt ctaaatacct cgcgcgtgtg ggagaa 516 SEQ ID NO: 9 atgatgcgcgtgtacgacta tcaaggaaga aagaggactt aatttcttac cttctaacca 60 ccatattctttttgctggat gcttgctcgt ctcgatgaca attgtgaacc tcttgtgtga 120 ccctgaccctgctgcaaggc tctccgaccg cacgcaaggc gcagccggcg cgtccggagg 180 cgatcggatccaatccagtc gtcctcccgc agcccgggca cgtttgccca tgcaggccct 240 tccacaccgctcaagagact cccgaacacc gcccactcgg cactcgcttc ggctgccgag 300 tgcgcgtttgagtttgccct gccacagaag acacc 335 SEQ ID NO: 10 atgatgcgcg tgtacgactatcaaggaaga aagaggactt aatttcttac cttctaacca 60 ccatattctt tttgctggatgcttgctcgt ctcgatgaca attgtgaacc tcttgtgtga 120 ccctgaccct gctgcaaggctctccgaccg cacgcaaggc gcagccggcg cgtccggagg 180 cgatcggatc caatccagtcgtcctcccgc agcccgggca cgtttgccca tgcaggccct 240 tccacaccgc tcaagagactcccgaacacc gcccactcgg cactcgcttc ggctgccgag 300 tgcgcgtttg agtttgccctgccacaggag acatc 335 SEQ ID NO: 11 cccgggcgag ctgtacgcct acggagcgaggcctggtgtg accgttgcga tctcgccagc 60 agacgtcgcg gagcctcgtc ccaaaggccctttctgatcg agcttgtcgt ccactggacg 120 ctttaagttg cgcgcgcgat gggataaccgagctgatctg cactcagatt ttggtttgtt 180 ttcgcgcatg gtgcagcgag gggaggtactacgctggggt acgagatcct ccggattccc 240 agaccgtgtt gccggcattt acccggtcatcgccagcgat tcgggacgac aaggccttat 300 cctgtgctga gacgctcgag cacgtttataaaattgtggg taccgcggta tgcacagcgt 360 tcaacacgcg ccacgccgaa attggttggtgggggagcac gtatgggact gacgtatggc 420 cagcagcgaa cactcaccga acaagtgccaatgtatacct tgcatcaatg atgctccggc 480 agcttcgatt gactgtctcg aaaaagtgtgagcaagcaga tcatgtggcc gctctgtcgc 540 gcagcacctg acgcattcga cacccacggcaatgcccagg ccagggaata gagagtaaga 600 caactcccat tgttcagcaa aacattgcactgcagtgcct tcacaactat acaatgaatg 660 ggagggaata tgggctctgc atgggacagcttagctggga cattcggcta ctgaacaaga 720 aaaccccacg agaaccaatt ggcgaaacctgccgggagga ggtgatcgtt tctgtaaatg 780 gcttacgcat tcccccccgg cggctcacgaggggtgtggt gaaccctgcc agctgatcaa 840 gtgcttgctg acgtcggcca gggaggtgtatgtgattggg ccgtggggcg tgagttatcc 900 taccgccgga cccgcgaagt cacatgacgaatggccgtgc gggatgacga gagcacgact 960 cgctctttct tcgccggccc ggcttcatggaggacaataa taaagggtgg ccaccggcaa 1020 cagccctcca tacctgaacc gattccagacccaaacctct tgaattttga gggatccagt 1080 tcaccggtat agtcacg 1097SEQ ID NO: 12 atccccgggc gagctgtacg cctacggagc gaggcctggt gtgaccgttgcgatctcgcc 60 agcagacgtc gcggagcctc gtcccaaagg ccctttctga tcgagcttgtcgtccactgg 120 acgctttaag ttgcgcgcgc gatgggataa ccgagctgat ctgcactcagattttggttt 180 gttttcgcgc atggtgcagc gaggggaggt actacgctgg ggtacgagatcctccggatt 240 cccagaccgt gttgccggca tttacccggt catcgccagc gattcgggacgacaaggcct 300 tatcctgtgc tgagacgctc gagcacgttt ataaaattgt ggtcaccgtggtacgcacag 360 cgtccaacac gcgccacgcc gaaattcgtt ggtgggggag cacgtatcggactgacgtat 420 ggccagcagc gaacactcac caaacaggtg ccaatgtata gcttgcatcaatgatgctct 480 ggcagcttcg attgactgtc tcgaaaaagt gtgtgcaaac agattatgtggccgctctgt 540 ggccgcgcag cacctgacgc actcgacacc cacggcaatg cccaggccaaggaacagaga 600 gtaagacaac tcccattgtt cagtaaaaca ttgcactgca gtgccttcacaaacatacaa 660 cgaatgggag ggaatatggg cttcgaatgg gacagcttag ctgggacattcggttactga 720 acaagaaaac cccacgagaa ccaactggcg aaacctgccg ggaggaggtgatcgtttttg 780 taaatggctt acgcattccc cccccggcgg ctcacggggg gtgtggtgaaccctgccagc 840 tgatcaagtg cttgctgacg tcggccaggg aggtgtatgt gatttggccgtggggcgtga 900 gttatcctac cgccggaccc gcgaagtcac atgacgaatg gccgtgcgggatgacgagag 960 cagggctcgc tctttcttcg ccggcccggc ttcatggagg acaataataaagggtggcca 1020 ccggcaacag ccctccatac ctgaaccga ttccagaccca aacctcttgaattttgaggg 1080 atccagttca ccggtatagt cacga 1105 SEQ ID NO: 13gcgagtggtt ttgctgccgg gaagggagtg gggagcgtcg agcgagggac gcggcgctcg 60aggcgcacgt cgtctgtcaa cgcgcgcggc cctcgcggcc cgcggcccca cccagctcta 120atcatcgaaa actaagaggc tccacacgcc tgtcgtagaa tgcatgggat tcgccagtag 180accacgatct gcgccgaaga agctggtcta cccgacgttt tttgttgctc ctttattctg 240aatgatatga agatagtgtg cgcagtgcca cgcataggca tcaggagcaa gggaggacgg 300gtcaacttga aagaaccaaa ccatccatcc gagaaatgcg catcatcttt gtagtaccat 360caaacgcctt ggccaatgtc ttctgcatgg acaacacaac ctgctcctgg ccacacggtc 420gacttggagc gccccatgcg cccaggtcgc cacgacccgc ggcccagcgc gcggcgattc 480gcctcacgag atcccggcgg acccggcacg cccgcgggcc gacggtgcgc ttggcgatgc 540tgctcattaa cccacggccg tcacccgatc cacatgctct ttttcaacac atccacattg 600gaatagagct ctaccagggt gagtactgca ttctttgggg ctgggaggac cccactcgac 660acctggtcct tcatcggccg aaagcccgaa cctgagcgct tccccgcccc gttcctcatc 720cccgactttc cgatggccca ttgcagtttc aaac 754 SEQ ID NO: 14 atctgggtggaggactggga gtaagatgta aggatattaa ttaaacattc tagtttgttg 60 atggcacaacagtcaatgca tttcagtcgt cttgctcctt ataacctatg cgtgtgccat 120 cgccggccatgcacctgtgg cgtggtaccg accatcgggg agaggcccga gattcggagg 180 tacctcccgccctgggcgag cccttcacgt gacggcacaa gtcccttgca tcggcccgcg 240 agcacggaatacagagcccc gtgcccccca cgggccctca catcatccac tccattgttc 300 ttgccacaccgatcagca 318 SEQ ID NO: 15 tgggtggagg actgggaaga agatgtaagg atatcaatttaacattctag tttgttgatg 60 gcacaacagt cactgaatac cgggcgtctg gctgctaaaatagccggagc gtgtgccatc 120 gccggccatg catctgtggc gtggtaccga ccatcagggagaggcccgag attcggaggt 180 acctcccgcc ctgggcgagc ccttcacgtg acggcacaagtcccttgcat cggcccgcga 240 gcacggaata cagagccccg tgctccccac gggccctcacatcatccact ccattgttct 300 tgccacaccg  atcagc 316 SEQ ID NO: 16ataacgaggc acaatgatcg atatttctat cgaacaactg tatttagccc tgtacgtacc 60ccgctcttgg gccagcccgt ccgtgcttgc cttcggaaaa ttgcatggcg cctcatgcaa 120actcgcgctc tcacagcaga tctcgcccag ctcccgggag agcaatcgcg ggtggggccc 180ggggcgaatc caggacgcgc cccgcggggc cgctccactc gccagggcca atgggcggct 240tatagtcctg gcatgggctc tgcatgcaca gtatcgcagt ttgggcgagg tgttgccccc 300gcgatttcga atacgcgacg cccggtactc gtgcgagaac agggttcttgPrototheca moriformis (UTEX 1435)Amt02 promoter SEQ ID NO: 17TCACCAGCGGACAAAGCACCGGTGTATCAGGTCCGTGTCATCCACTCTAAAGAGCTCGACTACGACCTACTGATGGCCCTAGATTCTTCATCAAAAACGCCTGAGACACTTGCCCAGGATTGAAACTCCCTGAAGGGACCACCAGGGGCCCTGAGTTGTTCCTTCCCCCCGTGGCGAGCTGCCAGCCAGGCTGTACCTGTGATCGGGGCTGGCGGGAAAACAGGCTTCGTGTGCTCAGGTTATGGGAGGTGCAGGACAGCTCATTAAACGCCAACAATCGCACAATTCATGGCAAGCTAATCAGTTATTTCCCATTAACGAGCTATAATTGTCCCAAAATTCTGGTCTACCGGGGGTGATCCTTCGTGTACGGGCCCTTCCCTCAACCCTAGGTATGCGCACATGCGGTCGCCGCGCAACGCGCGCGAGGGCCGAGGGTTTGGGACGGGCCGTCCCGAAATGCAGTTGCACCCGGATGCGTGGCACCTTTTTTGCGATAATTTATGCAATGGACTGCTCTGCAAAATTCTGGCTCTGTCGCCAACCCTAGGATCAGCGGTGTAGGATTTCGTAATCATTCGTCCTGATGGGGAGCTACCGACTGCCCTAGTATCAGCCCGACTGCCTGACGCCAGCGTCCACTTTTGTGCACACATTCCATTCGTGCCCAAGACATTTCATTGTGGTGCGAAGCGTCCCCAGTTACGCTCACCTGATCCCCAACCTCCTTATTGTTCTGTCGACAGAGTGGGCCCAGAGGCCGGTCGCAGCC Prototheca moriformis (UTEX 1435) Amt03 promoterSEQ ID NO: 18Ggccgacaggacgcgcgtcaaaggtgctggtcgtgtatgccctggccggcaggtcgttgctgctgctggttagtgattccgcaaccctgattttggcgtcttattttggcgtggcaaacgctggcgcccgcgagccgggccggcggcgatgcggtgccccacggctgccggaatccaagggaggcaagagcgcccgggtcagttgaagggctttacgcgcaaggtacagccgctcctgcaaggctgcgtggtggaattggacgtgcaggtcctgctgaagttcctccaccgcctcaccagcggacaaagcaccggtgtatcaggtccgtgtcatccactctaaagagctcgactacgacctactgatggccctagattcttcatcaaaaacgcctgagacacttgcccaggattgaaactccctgaagggaccaccaggggccctgagttgttccttccccccgtggcgagctgccagccaggctgtacctgtgatcgaggctggcgggaaaataggcttcgtgtgctcaggtcatgggaggtgcaggacagctcatgaaacgccaacaatcgcacaattcatgtcaagctaatcagctatttcctcttcacgagctgtaattgtcccaaaattctggtctaccgggggtgatccttcgtgtacgggcccttccctcaaccctaggtatgcgcgcatgcggtcgccgcgcaactcgcgcgagggccgagggtttgggacgggccgtcccgaaatgcagttgcacccggatgcgtggcaccttttttgcgataatttatgcaatggactgctctgcaaaattctggctctgtcgccaaccctaggatcagcggcgtaggatttcgtaatcattcgtcctgatggggagctaccgactaccctaatatcagcccgactgcctgacgccagcgtccacttttgtgcacacattccattcgtgcccaagacatttcattgtggtgcgaagcgtccccagttacgctcacctgtttcccgacctccttactgttctgtcgacagagcgggcccacaggccggtcgcagcc pSZ3840/D2554 transforming construct (CpauLPAAT1)SEQ ID NO: 19 gctcttccgctaacggaggtctgtcaccaaatggaccccgtctattgcgggaaaccacggcgatggcacgtttcaaaacttgatgaaatacaatattcagtatgtcgcgggcggcgacggcggggagctgatgtcgcgctgggtattgcttaatcgccagcttcgcccccgtcttggcgcgaggcgtgaacaagccgaccgatgtgcacgagcaaatcctgacactagaagggctgactcgcccggcacggctgaattacacaggcttgcaaaaataccagaatttgcacgcaccgtattcgcggtattttgttggacagtgaatagcgatgcggcaatggcttgtggcgttagaaggtgcgacgaaggtggtgccaccactgtgccagccagtcctggcggctcccagggccccgatcaagagccaggacatccaaactacccacagcatcaacgccccggcctatactcgaaccccacttgcactctgcaatggtatgggaaccacgggg

gcctgacgccccagatgggctgggacaactggaacacgttcgcctgcgacgtctccgagcagctgctgctggacaggccgaccgcatctccgacctgggcctgaaggacatgggctacaagtacatcatcctggacgactgctggtcctccggccgcgactccgacggcttcctggtcgccgacgagcagaagttccccaacggcatgggccacgtcgccgaccacctgcacaacaactccttcctgttcggcatgtactcctccgcgggcgagtacacgtgcgccggctaccccggctccctgggccgcgaggaggaggacgcccagacttcgcgaacaaccgcgtggactacctgaagtacgacaactgctacaacaagggccagttcggcacgcccgagatctcctaccaccgctacaaggccatgtccgacgccctgaacaagacgggccgccccatcttctactccctgtgcaactggggccaggacctgaccactactggggctccggcatcgcgaactcctggcgcatgtccggcgacgtcacggcggagttcacgcgccccgactcccgctgcccctgcgacggcgacgagtacgactgcaagtacgccggcttccactgctccatcatgaacatcctgaacaaggccgcccccatgggccagaacgcgggcgtcggcggctggaacgacctggacaacctggaggtcggcgtcggcaacctgacggacgacgaggagaaggcgcacttctccatgtgggccatggtgaagtcccccctgatcatcggcgcgaacgtgaacaacctgaaggcctcctcctactccatctactcccaggcgtccgtcatcgccatcaaccaggactccaacggcatccccgccacgcgcgtctggcgctactacgtgtccgacacggacgagtacggccagggcgagatccagatgtggtccggccccctggacaacggcgaccaggtcgtggcgctgctgaacggcggctccgtgtcccgccccatgaacacgaccctggaggagatcttcttcgactccaacctgggctccaagaagctgacctccacctgggacatctacgacctgtgggcgaaccgcgtcgacaactccacggcgtccgccatcctgggccgcaacaagaccgccaccggcatcctgtacaacgccaccgagcagtcctacaaggacggcctgtccaagaacgacacccgcctgttcggccagaagatcggctccctgtcccccaacgcgatcctgaacacgaccgtccccgcccacggcatcgcgttctaccgcctgcgcccc

acacttgctgccttgacctgtgaatatccctgccgcttttatcaaacagcctcagtgtgtttgatcttgtgtgtacgcgcttttgcgagttgctagctgcttgtgctatttgcgaataccacccccagcatccccttccctcgtttcatatcgcttgcatcccaaccgcaacttatctacgctgtcctgctatccctcagcgctgctcctgctcctgctcactgcccctcgcacagccttggtttgggctccgcctgtattctcctggtact

atcaacctgttccaggccctgtgcttcgtgctggtgtggcccctgtccaagaacgcctaccgccgcatcaaccgcgtgttcgccgagctgctgctgtccgagctgctgtgcctgttcgactggtgggccggcgccaagctgaagctgttcaccgaccccgagaccttccgcctgatgggcaaggagcacgccctggtgatcatcaaccacatgaccgagctggactggatgctgggctgggtgatgggccagcacctgggctgcctgggctccatcctgtccgtggccaagaagtccaccaagttcctgcccgtgctgggctggtccatgtggttctccgagtacctgtacatcgagcgctcctgggccaaggaccgcaccaccctgaagtcccacatcgagcgcctgaccgactaccccctgcccactggatggtgatcttcgtggagggcacccgcttcacccgcaccaagctgctggccgcccagcagtacgccgcctcctccggcctgcccgtgccccgcaacgtgctgatcccccgcaccaagggcttcgtgtcctgcgtgtcccacatgcgctccttcgtgcccgccgtgtacgacgtgaccgtggccttccccaagacctcccccccccccaccctgctgaacctgttcgagggccagtccatcgtgctgcacgtgcacatcaagcgccacgccatgaaggacctgcccgagtccgacgacgccgtggcccagtggtgccgcgacaagttcgtggagaaggacgccctgctggacaagcacaacgccgaggacaccttctccggccaggaggtgcaccgcaccggctcccgccccatcaagtccctgctggtggtgatctcctgggtggtggtgatcaccttcggcgccctgaagttcctgcagtggtcctcctggaagggcaaggccttctccgtgatcggcctgggcatcgtgaccctgctgatgcacatgctgatcctgtcctcccaggccgagcgctc

gacctgtgaatatccctgccgcttttatcaaacagcctcagtgtgtttgatcttgtgtgtacgcgcttttgcgagttgctagctgcttgtgctatttgcgaataccacccccagcatccccttccctcgtttcatatcgcttgcatcccaaccgcaacttatctacgctgtcctgctatccctcagcgctgctcctgctcctgctcactgcccctcgcacagccttggtttgggctccgcctgtattctcctggtactgcaacctgtaaaccagcactgcaatgctgatgcacgggaagtagtgggatgggaacacaaatggaaagctt gagctcagcggcgacggtcctgctaccgtacgacgttgggcacgcccatgaaagtttgtataccgagcttgttgagcgaactgcaagcgcggctcaaggatacttgaactcctggattgatatcggtccaataatggatggaaaatccgaacctcgtgcaagaactgagcaaacctcgttacatggatgcacagtcgccagtccaatgaacattgaagtgagcgaactgttcgcttcggtggcagtactactcaaagaatgagctgctgttaaaaatgcactctcgttctctcaagtgagtggcagatgagtgctcacgccttgcacttcgctgcccgtgtcatgccctgcgccccaaaatttgaaaaaagggatgagattattgggcaatggacgacgtcgtcgctccgggagtcaggaccggcggaaaataagaggcaacacactccgcttctta gctcttc pSZ3841/D2555 (CpaiLPAAT1) SEQ ID NO: 20

gccttctgcttcgtgctgatctcccccctgtccaagaacgcctaccgccgcatcaaccgcgtgacgccgagctgctgcccctggagacctgtggctgttccactggtgcgccggcgccaagctgaagctgttcaccgaccccgagaccttccgcctgatgggcaaggagcacgccctggtgatcatcaaccacaagatcgagctggactggatggtgggctgggtgctgggccagcacctgggctgcctgggctccatcctgtccgtggccaagaagtccaccaagttcctgcccgtgttcggctggtccctgtggttctccggctacctgttcctggagcgctcctgggccaaggacaagatcaccctgaagtcccacatcgagtccctgaaggactaccccctgcccttctggctgatcatcttcgtggagggcacccgcttcacccgcaccaagctgctggccgcccagcagtacgccgcctcctccggcctgcccgtgccccgcaacgtgctgatcccccacaccaagggcttcgtgtcctccgtgtcccacatgcgctccttcgtgcccgccatctacgacgtgaccgtggccttccccaagacctcccccccccccaccatgctgaagctgacgagggccagtccgtggagctgcacgtgcacatcaagcgccacgccatgaaggacctgcccgagtccgacgacgccgtggcccagtggtgccgcgacaagttcgtggagaaggacgccctgctggacaagcacaactccgaggacaccttctccggccaggaggtgcaccacgtgggccgccccatcaaggccctgctggtggtgatctcctgggtggtggtgatcatcttcggcgccctgaagttcctgctgtggtcctccctgctgtcctcctggaagggcaaggccactccgtgatcggcctgggcatcgtggccggcatcgtgaccctgctgatgcacatcctgatcctgtcctcccaggccgaggg

pSZ3842/D2556(CigneaLPAAT1) SEQ ID NO: 21

gccctgtgcttcgtgctgatctggcccctgtccaagaacgtgtaccgccgcatcaaccgcgtgacgccgagctgctgctgatggacctgctgtgcctgttccactggtgggccggcgccaagatcaagctgacaccgaccccgagaccttccgcctgatgggcatggagcacgccctggtgatcatgaaccacaagaccgacctggactggatggtgggctggatcctgggccagcacctgggctgcctgggctccatcctgtccatcgccaagaagtccaccaagttcatccccgtgctgggctggtccgtgtggactccgagtacctgttcctggagcgctcctgggccaaggacaagtccaccctgaagtcccacatggagaagctgaaggactaccccctgcccttctggctggtgatcttcgtggagggcacccgcttcacccgcaccaagctgctggccgcccagcagtacgccgcctcctccggcctgcccgtgccccgcaacgtgctgatcccccacaccaagggcttcgtgtcctgcgtgtccaacatgcgctccacgtgcccgccgtgtacgacgtgaccgtggccttccccaagtcctcccccccccccaccatgctgaagctgttcgagggccagtccatcgtgctgcacgtgcacatcaagcgccacgccctgaaggacctgcccgagtccgacgacgccgtggcccagtggtgccgcgacaagttcgtggagaaggacgccctgctggacaagcacaacgccgaggacaccttctccggccaggaggtgcaccacatcggccgccccatcaagtccctgctggtggtgatcgcctgggtggtggtgatcatcttcggcgccctgaagttcctgcagtggtcctccctgctgtccacctggaagggcaaggccttctccgtgatcggcctgggcatcgccaccctgctgatgcacatgctgatcctgtcctcccaggccgagcgctccaacc

pSZ3844/D2557 (ChookLPAAT1) SEQ ID NO: 22

gccttctgcttcgtgctgatctcccccctgtccaagaacgcctaccgccgcatcaaccgcgtgacgccgagctgctgcccctggagacctgtggctgttccactggtgcgccggcgccaagctgaagctgttcaccgaccccgagaccttccgcctgatgggcaaggagcacgccctggtgatcatcaaccacaagatcgagctggactggatggtgggctgggtgctgggccagcacctgggctgcctgggctccatcctgtccgtggccaagaagtccaccaagttcctgcccgtgttcggctggtccctgtggttctccgagtacctgttcctggagcgctcctgggccaaggacaagatcaccctgaagtcccacatcgagtccctgaaggactaccccctgcccttctggctgatcatcttcgtggagggcacccgcttcacccgcaccaagctgctggccgcccagcagtacgccgcctcctccggcctgcccgtgccccgcaacgtgctgatcccccacaccaagggcttcgtgtcctccgtgtcccacatgcgctccttcgtgcccgccatctacgacgtgaccgtggccttccccaagacctcccccccccccaccatgctgaagctgacgagggccagtccgtggagctgcacgtgcacatcaagcgccacgccatgaaggacctgcccgagtccgacgacgccgtggcccagtggtgccgcgacaagttcgtggagaaggacgccctgctggacaagcacaactccgaggacaccttctccggccaggaggtgcaccacgtgggccgccccatcaaggccctgctggtggtgatctcctgggtggtggtgatcatcttcggcgccctgaagttcctgctgtggtcctccctgctgtcctcctggaagggcaaggccactccgtgatcggcctgggcatcgtggccggcatcgtgaccctgctgatgcacatcctgatcctgtcctcccaggccgaggg

CpauLPAAT1 SEQ ID NO: 23MAIPAAAVIFLFGLLFFTSGLIINLFQALCFVLVWPLSKNAYRRINRVFAELLLSELLCLFDWWAGAKLKLFTDPETFRLMGKEHALVIINHMTELDWMLGWVMGQHLGCLGSILSVAKKSTKFLPVLGWSMWFSEYLYIERSWAKDRTTLKSHIERLTDYPLPFWMVIFVEGTRFTRTKLLAAQQYAASSGLPVPRNVLIPRTKGFVSCVSHMRSFVPAVYDVTVAFPKTSPPPTLLNLFEGQSIVLHVHIKRHAMKDLPESDDAVAQWCRDKFVEKDALLDKHNAEDTFSGQEVHRTGSRPIKSLLVVISWVVVITFGALKFLQWSSWKGKAFSVIGLGIVTLLMHMLILSSQAERSSNPAKVAQAKLKTELSISKKATDKEN SEQ ID NO: CprocL2AAT1 SEQ ID NO: 24MAIPAAAVIFLFGLIFFASGLIINLFQALCFVLIWPISKNAYRRINRVFAELLLSELLCLFDWWAGAKLKLFTDPETFRLMGKEHALVIINHMTELDWMVGWVMGQHFGCLGSILSVAKKSTKFLPVLGWSMWFTEYLYIERSWNKDKSTLKSHIERLKDYPLPFWLVIFAEGTRFTQTKLLAAQQYAASSGLPVPRNVLIPRTKGFVSCVSHMRSFVPAVYDLTVAFPKTSPPPTLLNLFEGQSVVLHVHIKRHAMKDLPESDDEVAQWCRDKEVEKDALLDKHNAEDTFSGQELQHTGRRPIKSLLVVISWVVVIAFGALKFLQWSSWKGKAFSVIGLGIVTLLMHMLILSSQAERSKPAKVAQAKLKTELSISKTVTDKEN CprocL2AAT1b SEQ ID NO: 25MAIPAAAVIFLFGLIFFASGLIINLFQALCFVLIWPISKNAYRRINRVFAELLLSELLCLFDWWAGAKLKLFTDPETFRLMGKEHALVIINHMTELDWMVGWVMGQHFGCLGSILSVAKKSTKFLPVLGWSMWFTEYLYIERSWNKDKSTLKSHIERLKDYPLPFWLVIFAEGTRFTQTKLLAAQQYAASSGLPVPRNVLIPRTKGFVSCVSHMRSFVPAVYDLTVAFPKTSPPPTLLNLFEGQSVVLHVHIKRHAMKDLPESDDEVAQWCRDKEVEK SEQ ID NO:CprocL2AAT2a SEQ ID NO: 26IVNLVQAVCFVLVRPLSKNTYRRINRVVAELLWLELVWLIDWWAGVKIKVFTDHETFHLMGKEHALVICNHKSDIDWLVGWVLAQRSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYLFLERNWAKDESTLKSGLNRLKDYPLPFWLALFVEGTRFTRAKLLAAQQYAASSGLPVPRNVLIPRTKGFVSSVSHMRSFVPAIYDVTVAIPKTSPPPTLIRMFKGQSSVLHVHLKRHVMKDLPESDDAVAQWCRDIFVEKDALLDKHNADDTFSGQELQDTGRPIKSLLVVISWAVLEVFGAVKFLQWSSLLSSWKGLAFSGIGLGIITLLMHILILFSQSERSTPAKVAPAKAKIEGESSKTEMEKEKCprocL2AAT2b SEQ ID NO: 27IVNLVQAVCFVLVRPLSKNTYRRINRVVAELLWLELVWLIDWWAGVKIKVFTDHETFHLMGKEHALVICNHKSDIDWLVGWVLAQRSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYLFLERNWAKDESTLKSGLNRLKDYPLPFWLALFVEGTRFTRAKLLAAQQYAASSGLPVPRNVLIPRTKGFVSSVSHMRSFVPAIYDVTVAIPKTSPPPTLIRMFKGQSSVLHVHLKRHVMKDLPESDDAVAQWCRDIFVEKDALLDKHNADDTFSGQELQDTGRPIKSLLVCpaiLPAAT1 SEQ ID NO: 28MAIPSAAVVFLFGLLFFTSGLIINLFQAFCFVLISPLSKNAYRRINRVFAELLPLEFLWLFHWCAGAKLKLFTDPETFRLMGKEHALVIINHKIELDWMVGWVLGQHLGCLGSILSVAKKSTKFLPVFGWSLWFSGYLFLERSWAKDKITLKSHIESLKDYPLPFWLIIFVEGTRFTRTKLLAAQQYAASSGLPVPRNVLIPHTKGFVSSVSHMRSFVPAIYDVTVAFPKTSPPPTMLKLFEGQSVELHVHIKRHAMKDLPESDDAVAQWCRDKEVEKDALLDKHNSEDTFSGQEVHHVGRPIKALLVVISWVVVIIFGALKFLLWSSLLSSWKGKAFSVIGLGIVAGIVTLLMHILILSSQAEGSNPVKAAPAKLKTELSSSKKVTNKEN ChookLPAAT1 SEQ ID NO: 29MAIPSAAVVFLFGLLFFTSGLIINLFQAFCFVLISPLSKNAYRRINRVFAELLPLEFLWLFHWCAGAKLKLFTDPETFRLMGKEHALVIINHKIELDWMVGWVLGQHLGCLGSILSVAKKSTKFLPVFGWSLWFSEYLFLERSWAKDKITLKSHIESLKDYPLPFWLIIFVEGTRFTRTKLLAAQQYAASSGLPVPRNVLIPHTKGFVSSVSHMRSFVPAIYDVTVAFPKTSPPPTMLKLFEGQSVELHVHIKRHAMKDLPESDDAVAQWCRDKEVEKDALLDKHNSEDTFSGQEVHHVGRPIKALLVVISWVVVIIFGALKFLLWSSLLSSWKGKAFSVIGLGIVAGIVTLLMHILILSSQAEGSNPVKAAPAKLKTELSSSKKVTNKEN ChookL2AAT2a SEQ ID NO: 30LSLLFFVSGLIVNLVQAVCFVLIRPLSKNTYRRINRVVAELLWLELVWLIDWWAGVKIKVFTDHETFNLMGKEHALVVCNHKSDIDWLVGWVLAQRSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYLFLERSWAKDESTLKSGLKRLKDYPLPFWLALFVEGTRFTQAKLLAAQQYAASSGLPVPRNVLIPRTKGFVSSVSHMRSFVPAIYDVTVAIPKTSVPPTMLRIFKGQSSVLHVHLKRHLMKDLPESDDAVAQWCRDIFVEKDALLDKHNAEDTFSGQELQDIGRPIKSLLVVISWAVLVIFGAVKFLQWSSLLSSWKGLAFSGIGLGIVTLLMHILILFSQSERSTPAKVAPAKPKNEGESSKTEMEKEH ChookL2AAT2b SEQ ID NO: 31QIKVFTDHETFNLMGKEHALVVCNHKSDIDWLVGWVLAQWSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYLFLERSWAKDESTLKSGLKRLKDYPLPFWLALFVEGTRFTQAKLLAAQQYAASSGLPVPRNVLIPRTKGFVSSVSHMRSFVPAIYDVTVAIPKTSVPPTMLRIFKGQSSVLHVHLKRHLMKDLPESDDAVAQWCRDIFVEKDALLDKHNAEDTFSGQELQDIGRPIKSLLVVISWAVLVIFGAVKFLQWSSLLSSWKGLAFSGIGLGIVTLLMHILILFSQSERSTPAKVAPAKLKKEGESSKPETDKQN ChookL2AAT3a SEQ ID NO: 32LSLLFFVSGLIVNLVQAVCFVLIRPLLKNTYRRINRVVAELLWLELVWLIDWWAGIKIKVFTDHETFHLMGKEHALVICNHKSDIDWLVGWVLAQRSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYLFLERNWAKDESTLKSGLNRLKDYPLPFWLALFVEGTRFTRAKLLAAQQYAASSGLPVPRNVLIPRTKGFVSSVSQMRSFVPAIYDVTVAIPKTSPPPTLLRMFKGQSSVLHVHLKRHLMNDLPESDDAVAQWCRDIFVEKDALLDKHNAEDTFSGQELQDTGRPIKSLLVVISWATLVVFGAVKFLQWSSLLSSWKGLAFSGIGLGIITLLMHILILFSQSERSTPAKVAPAKPKNEGESSKTEMEKEH ChookL2AAT3b SEQ ID NO: 33LSLLFFVSGLIVNLVQAVCFVLIRPLLKNTYRRINRVVAELLWLELVWLIDWWAGIKIKVFTDHETFHLMGKEHALVICNHKSDIDWLVGWVLAQRSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYLFLERNWAKDESTLKSGLNRLKDYPLPFWLALFVEGTRFTRAKLLAAQQYAASSGLPVPRNVLIPRTKGFVSSVSQMRSFVPAIYDVTVAIPKTSPPPTLLRMFKGQSSVLHVHLKRHLMNDLPESDDAVAQWCRDIFVEKDALLDKHNAEDTFSGQELQDIGRPIKSLLVVISWAVLEIFGAVKFLQWSSLLSSWKGLAFSGIGLGIVTLLMHILILFSQSERSTPAKVAPAKPKKEGESSKPETDKEN CigneaLPAAT1 SEQ ID NO: 34MAIAAAAVIFLFGLLFFASGIIINLFQALCFVLIWPLSKNVYRRINRVFAELLLMDLLCLFHWWAGAKIKLFTDPETFRLMGMEHALVIMNHKTDLDWMVGWILGQHLGCLGSILSIAKKSTKFIPVLGWSVWFSEYLFLERSWAKDKSTLKSHMEKLKDYPLPFWLVIFVEGTRFTRTKLLAAQQYAASSGLPVPRNVLIPHTKGFVSCVSNMRSFVPAVYDVTVAFPKSSPPPTMLKLFEGQSIVLHVHIKRHALKDLPESDDAVAQWCRDKEVEKDALLDKHNAEDTFSGQEVHHIGRPIKSLLVVIAWVVVIIFGALKFLQWSSLLSTWKGKAFSVIGLGIATLLMHMLILSSQAERSNPAKVAKCigneaLPAAT2 SEQ ID NO: 35MAIAAAAVIFLFGLLFFASGIIINLFQALCFVLIWPLSKNVYRRINRVFAELLLMDLLCLFHWWAGAKIKLFTDPETFRLMGMEHALVIMNHKTDLDWMVGWILGQHLGCLGSILSIAKKSTKFIPVLGWSVWFSEYLFLERSWAKDESTLKSGLNRLKDYPLPFWLALFVEGTRFTRAKLLAAQQYAASSGLPVPKNVLIPRTKGFVSSVSHMRSFVPAIYDVTVAIPKTSAPPTLLRMFKGQSSVLHVHLKRHLMKDLPESDDAVAQWCRDIFVEKDALLDKHNAEDTFSGQELHDIGRPVKSLLVVISWAMLVVFGAVKFLQWSSLLSSWKGLAFSGIGLGIITLLMHILILFSQSERSTPAKVAPAKQKNNEGESSKTEMEKEH DcLPAAT1 SEQ ID NO: 36SGLVVNLIQAFFEVLVRPFSKNAYRKINRVVAELLWLELIWLIDWWAGVKIQLYTDPETFKLMGKEHALVICNHKSDIDWLVGWILAQRSGCLGSALAVMKKSSKFLPVIGWSMWFSEYLFLERSWAKDENTLKSGFQRLRDFPHAFWLALEVEGTRFTQAKLLAAQEYASSMGLPAPRNVLIPRTKGFVTAVTHMRPFVPAVYDVTLAIPKTSPPPTMLRLFKGQSSVVHIHLKRHLMSDLPKSDDSVAQWCKDAFVVKDNLLDKHKENDSFGDGVLQDTGRPLNSLVVVISWACLLIFGALKFFQWSSILSSWKGLAFSAVGLGIVTVLMQILIQFSQSERSNRPMPSKHAK DcLPAAT2SEQ ID NO: 37MAIPTAAYVVPLGAIFFFSGLLVNLIQAFFFITVWPLSKKTYIRINKVIVELLWLEFVWLADWWAGLKIEVYADAETFQLMGKEHALVICNHKSDIDWLVGWILAQRAGCLGSSFAVTKKSARYLPVVGWSIWFSGAIFLERSWEKDENTLKAGFQRLREFPCAFWLGLFVEGTRFTQAKLLAAQEYASTMGLPFPRNVLIPRTKGFIAAVNHMREFVPAIYDLTFAFPKDSPPPTMLRLLKGQPSVVHVHIKRHLMKDLPEKNEAVAQWCKDVFLVKDKLLDKHKDDGSFGDGELHEIGRPLKSLVVVTTWACLLILGTLKFLLWSSLLSSWKGLIFSATGLAVLTVLMQFLIQSTQSERSNPASLSKCcrLPAATla SEQ ID NO: 38LGLLFFISGLAVNLIQAVCFVFLRPLSKNTYRKINRVLAELLWLQLVWLVDWWAGVKIKVFADRESFNLMGKEHALVICNHKSDIDWLVGWVLAQRSGCLGSSLAVMKKSSKFLPVIGWSMWFSEYLFLERSWAKDESTLKEGLRRLKDFPRPFWLALFVEGTRFTQAKLLAAQEYATSQGLPVPRNVLIPRTKVHVHVKRHLMKELPETDEAVAQWCKDLFVEKDKLLDKHVAEDTFSDQPLQDIGRPVKPLLVVSSWACLVAYGALKFLQWSSLLSSWKGIAVSAVALAIVTILMQIMILFSQSERSIPAKVA CcrLPAAT1b SEQ ID NO: 39LGLLFFISGLAVNLIQAVCFVFLRPLSKNTYRKINRVLAELLWLQLVWLVDWWAGVKIKVFADRESFNLMGKEHALVICNHKSDIDWLVGWVLAQRSGCLGSSLAVMKKSSKFLPVIGWSMWFSEYLFLERSWAKDESTLKEGLRRLKDFPRPFWLALFVEGTRFTQAKLLAAQEYATSQGLPVPRNVLIPRTKGFVSAVSHMRSFVPAVYDMTVAIPKSSPSPTMLRLFKGQSSVVHVHVKRHLMKELPETDEAVAQWCKDLFVEKDKLLDKHVAEDTFSDQPLQDIGRPVKPLLVVSSWACLVAYGALKFLQWSSLLSSWKGIAVSAVALAIVTILMQIMILFSQSERSIPTKVA CcrL2AAT2aSEQ ID NO: 40MAIAAAAVVFLFGLLFFTSGLIINLAQAVCFVLIWPLSKNAYRRINRVFAELLLLELLWLFHWRAGAKLKLFADPETFRLFGKEHALVICNHRTDLDWMVGWVLGQHFGCLGSILSVAKKSTKFLPVLGWSMWFSEYLFLERSWAKDKSTLKSHTERLKDYPLPFWLGIFVEGTRFTRAKLLAAQQYAASSGLPVPRNVLIPHTKLHVHIKRYAMKDLPESDDAVAQWCRDIYVEKDAFLDKHNAEDTFSGQEVHHIGRPIKSLLVVISWVVVIIFGALKFLRWSSLLSSWKGKAFSVIGLGIVTLLVNILILSSQAERSNPAKVAPAKLKTELSPSKKVTNKEN CcrL2AAT2b SEQ ID NO: 41MAIAAAAVVFLFGLLFFTSGLIINLAQAVCFVLIWPLSKNAYRRINRVFAELLLLELLWLFHWRAGAKLKLFADPETFRLFGKEHALVICNHRTDLDWMVGWVLGQHFGCLGSILSVAKKSTKFLPVLGWSMWFSEYLFLERSWAKDKSTLKSHTERLKDYPLPFWLGIFVEGTRFTRAKLLAAQQYAASSGLPVPRNVLIPHTKGFVSSMSHMRSFVPAVYDLTVAFPKTSPPPTLLKLFEGQSVVLHVHIKRYAMKDLPESDDAVAQWCRDIYVEKDAFLDKHNAEDTFSGQEVHHIGRPIKSLLVVISWVVVIIFGALKFLRWSSLLSSWKGKAFSVIGLGIVTLLVNILILSSQAERSNPAKVAPAKLKTELSPSKKVTNKEN BrLPAATla SEQ ID NO: 42AAAVIVPLGILFFISGLVVNLLQAICYVLIRPLSKNTYRKINRVVAETLWLELVWIVDWWAGVKIQVFADNETFNRMGKEHALVVCNHRSDIDWLVGWILAQRSGCLGSALAVMKKSSKFLPVIGWSMWFSEYLFLERNWAKDESTLKSGLQRLNDFPRPFWLALFVEGTRFTEAKLKAAQEYAASSELPVPRNVLIPRTKGFVSAVSNMRSFVPAIYDMTVAIPKTSPPPTMLRLFKGQPSVVHVHIKCHSMKDLPESDDAIAQWCRDQFVAKDALLDKHIAADTFPGQQEQNIGRPIKSLAVVLSWSCLLILGAMKFLHWSNLFSSWKGIAFSALGLGIITLCMQILIRSSQSERSTPAKVVPAKPKDNHNDSGSSSQTE BrLPAAT1b SEQ ID NO: 43AAAVIVPLGILFFISGLVVNLLQAVCYVLVRPMSKNTYRKINRVVAETLWLELVWIVDWWAGVKIQVFADDETFNRMGKEHALVVCNHRSDIDWLVGWILAQRSGCLGSALAVMKKSSKFLPVIGWSMWFSEYLFLERNWAKDESTLKSGLQRLNDFPRPFWLALFVEGTRFTEAKLKAAQEYAASSELPVPRNVLIPRTKGFVSAVSNMRSFVPAIYDMTVAIPKTSPPPTMLRLFKGQPSVVHVHIKCHSMKDLPESDDAIAQWCRDQFVAKDALLDKHIAADTFPGQQEQNIGRPIKSLAVVLSWSCLLILGAMKFLHWSNLFSSWKGIAFSALGLGIITLCMQILIRSSQSERSTPAKVVPAKPKDNHNDSGSSSQTE BrLPAAT1c SEQ ID NO: 44MAIAAAVIVPLGLLFFISGLLMNLLQAICYVLVRPLSKNTYRKINRVVAETLWLELVWIVDWWAGVKIKVFADNETFSRMGKEHALVVCNHRSDIDWLVGWILAQRSGCLGSALAVMKKSSKFLPVIGWSMWFSEYLFLERNWAKDESTLKSGLQRLNDFPRPFWLALFVEGTRFTEAKLKAAQEYAASSELPVPRNVLIPRTKGFVSAVSNMRSFVPAIYDMTVAIPKTSPPPTMLRLFKGQPSVVHVHIKCHSMKDLPESDDAIAQWCRDQFVAKDALLDKHIAADTFPGQQEQNIGRPIKSLAVVLSWSCLLILGAMKFLHWSNLFSSWKGIAFSALGLGIITLCMQILIRSSQSERSTPAKVVPAKPKDNHNDSGSSSQTE BjLPAAT1a SEQ ID NO: 45INLVVAETLWLELVWIVDWWAGVKIQVFADDETFNRMGKEHALVVCNHRSDIDWLVGWILAQRSGCLGSALAVMKKSSKFLPVIGWSMWFSEYLFLERNWAKDESTLKSGLQRLNDFPRPFWLALFVEGTRFTEAKLKAAQEYAASSELPVPRNVLIPRTKGFVSAVSNMRSFVPAIYDMTVAIPKTSPPPTMLRLFKGQPSVVHVHIKCHSMKDLPESDDAIAQWCRDQFVAKDALLDKHIAADTFPGQKEQNIGRPIKSLAVSLIKTFPWLHPHQLTNIFVLFQVVVSWACLLTLGAMKFLHWSNLFSSWKGIALSAFGLGIITLCMQILIRSSQSERSTPAKVAPAKPK BjLPAAT1bSEQ ID NO: 46INLVVAETLWLELVWIVDWWAGVKIQVFADDETFNRMGKEHALVVCNHRSDIDWLVGWILAQRSGCLGSALAVMKKSSKFLPVIGWSMWFSEYLFLERNWAKDESTLKSGLQRLNDFPRPFWLALFVEGTRFTEAKLKAAQEYAASSELPVPRNVLIPRTKGFVSAVSNMRSFVPAIYDMTVAIPKTSPPPTMLRLFKGQPSVVHVHIKCHSMKDLPEPEDEIAQWCRDQFVAKDALLDKHIAADTFPGQKEQNIGRPIKSLAVVVSWACLLTLGAMKFLHWSNLFSSWKGIALSAFGLGIITLCMQILIRSSQSERSTPAKVAPAKPK BjLPAAT1c SEQ ID NO: 47INLVVAETLWLELVWIVDWWAGVKIQVFADDETFNRMGKEHALVVCNHRSDIDWLVGWILAQRSGCLGSALAVMKKSSKFLPVIGWSMWFSEYLFLERNWAKDESTLKSGLQRLNDFPRPFWLALFVEGTRFTEAKLKAAQEYAASSELPVPRNVLIPRTKGFVSAVSNMRSFVPAIYDMTVAIPKTSPPPTMLRLFKGQPSVVHVHIKCHSMKDLPESDDAIAQWCRDQFVAKDALLDKHIAADTFPGQQEQNIGRPIKSLAVVLSWSCLLILGAMKFLHWSNLFSSWKGIAFSALGLGIITLCMQILIRSSQSERSTPAKVVPAKPKDNHNDSGSSSQTE BjLPAAT1d SEQ ID NO: 48INLVVAETLWLELVWIVDWWAGVKIQVFADDETFNRMGKEHALVVCNHRSDIDWLVGWILAQRSGCLGSALAVMKKSSKFLPVIGWSMWFSEYLFLERNWAKDESTLKSGLQRLNDFPRPFWLALFVEGTRFTEAKLKAAQEYAASSELPVPRNVLIPRTKGFVSAVSNMRSFVPAIYDMTVAIPKTSPPPTMLRLFKGQPSVVHVHIKCHSMKDLPESDDAIAQWCRDQFVAKDALLDKHIAADTFPGQQEQNIGRPIKSLAVSLS CcLPAAT1a SEQ ID NO: 49MAIGVAAIVVPLGLLFILSGLMVNLIQAICFILVRPLSKNMYRRVNRVVVELLWLELIWLIDWWGGVKVDVYADSETFQSLGKEHALVVSNHRSDIDWLVGWVLAQRSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYVFLERSWAKDESTLKSGLRRLKDFPRPFWLALFVEGTRFTQAKLLAAREYAASTGLPIPRNVLIPRTKGFVSAVSNMRSFVPAIYDVTVAIPKTQPSPTMLRIFNRQPSVVHVHIKRHSMNQLPQTDEGVGQWCKDIFVAKDALLDRHLAECcLPAAT1b SEQ ID NO: 50MAIGVAAIVVPLGLLFILSGLMVNLIQAICFILVRPLSKNMYRRVNRVVVELLWLELIWLIDWWGGVKVDVYADSETFQSLGKEHALVVSNHRSDIDWLVGWVLAQRSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYVFLERSWAKDESTLKSGLRRLKDFPRPFWLALFVEGTRFTQAKLLAAREYAASTGLPIPRNVLIPRTKGFVSAVSNMRSFVPAIYDVTVAIPKTQPSPTMLRIFNRQPSVVHVHIKRHSMNQLPQTDEGVAQWCKDIFVAKDALLDRHLAEGKFDEKEFKRIRRPIKSLLVISSWSFLLMFGVFKFLKWSALLSTWKGVAVSTTVLLLVTVVMYMFILFSQSERSSPRKVAPSGPENGUcLPAAT1a SEQ ID NO: 51MAIGVAAIVVPLGLLFILSGLIINLIQAICFILVRPLSKNMYRKVNRVVVELLWLELIWLIDWWGGVKVDVYADSETFQSLGKEHALVVSNHRSDIDWLVGWVLAQRSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYVFLERSWAKDESTLKSGLQRLKDFPRPFWLALFVEGTRFTQAKLLAAQEYAASTGLPIPRNVLIPRTKGEVSAVSNMRSFVPAIYDVTVAIPKTQPSPTMLRIFNRQPSVVHVHIKRHSMNQLPQTDEGVAQWCKDIFVAKDALLDRHLAEGKFDEKEFKLIRRPIKSLLVISSWSFLLMFGVFKFLKWSALLSTWKGVAVSTAVLLLVTVVMYMFILFSQSERSSPRKVAPIGPENGUcLPAAT1b SEQ ID NO: 52MAIGVAAIVVPLGLLFILSGLIINLIQAICFILVRPLSKNMYRKVNRVVVELLWLELIWLIDWWGGVKVDVYADSETFQSLGKEHALVVSNHRSDIDWLVGWVLAQRSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYVFLERSWAKDESTLKSGLQRLKDFPRPFWLALFVEGTRFTQAKLLAAQEYAASTGLPIPRNVLIPRTKGEVSAVSNMRSFVPAIYDVTVAIPKTQPSPTMLRIFNRQPSVVHVHIKRHSMNQLPQTDEGVAQWCKDIFVAKDALLDRHLAE LdLPAAT1SEQ ID NO: 53SLLFFMSGLVVNFIQAVFYVLVRPISKNTYRRINTLVAELLWLELVWVIDWWAGVKVQLYTDTESFRLMGKEHALLICNHRSDIDWLIGWVLAQRCGCLSSSIAVMKKSSKFLPVIGWSMWFSEYLFLERNWAKDENTLKSGLQRLNDFPKPFWLALFVEGTRFTKAKLLAAQEYAASAGLPVPRNVLIPRTKGFVSAVSNMRSFVPAIYDLTVAIPKTTEQPTMLRLFRGKSSVVHVHLKRHLMKDLPKTDDGVAQWCKDQFISKDALLDKHVAEDTFSGLEVQDIGRPMKSLVVVVSWMCLLCLGLVKFLQWSALLSSWKGMMITTFVLGIVTVLMHILIRSSQSEHSTPAK CaequLPAAT1aSEQ ID NO: 54QRSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYLFLERSWAKDESTLKSGLKRLKDYPLPFWLALFVEGTRFTQAKLLAAQQYAASSGLPVPRNVLIPRTKGEVSSVSHMRSFVPAIYDVTVAIPKMSTPPTMLRIFKGQSSVLHVHLKRHLMKDLPESDDAVAQWCRDIFVEKDALLDKHNAEDTFSGQELQDIGRPVKSLLVVISWAVLVIFGAVKFLQWSSLLSSWKGLAFSGIGLGIVTLLMHILILFSQSERSTPAKVAPAKPKKEGESSKTETEKEN CaequLPAAT1bSEQ ID NO: 55DWWAGVKIKVFTDHETLSLMGKEHALVISNHKSDIDWLVGWVLAQRSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYLFLERSWAKDESTLKSGLKRLKDYPLPFWLALFVEGTRFTQAKLLAAQQYAASSGLPVPRNVLIPRTKGEVSSVSHMRSFVPAIYDVTVAIPKMSTPPTMLRIFKGQSSVLHVHLKRHLMKDLPESDDAVAQWCRDIFVEKDALLDKHNAEDTFSGQELQDIGRPVKSLLV CaequLPAAT1c SEQ ID NO: 56DWWAGVKIKVFTDHETLSLMGKEHALVISNHKSDIDWLVGWVLAQRSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYLFLERSWAKDESTLKSGLKRLKDYPLPFWLALFVEGTRFTQAKLLAAQQYAASSGLPVPRNVLIPRTKGEVSSVSHMRSFVPAIYDVTVAIPKMSTPPTMLRIFKGQSSVLHVHLKRHLMKDLPESDDAVAQWCRDIFVEKDALLDKHNAEDTFSGQELQDIGRPVKSLLVVISWAVLVIFGAVKFLQWSSLLSSWKGLAFSGIGLGIVTLLMHILILFSQSERSTPAKVAPAKPKKEGESSKTETEKEN CaequLPAAT1d SEQ ID NO: 57QRSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYLFLERSWAKDESTLKSGLKRLKDYPLPFWLALFVEGTRFTQAKLLAAQQYAASSGLPVPRNVLIPRTKGEVSSVSHMRSFVPAIYDVTVAIPKMSTPPTMLRIFKGQSSVLHVHLKRHLMKDLPESDDAVAQWCRDIFVEKDALLDKHNAEDTFSGQELQDIGRPVKSLLV CglutLPAAT1aSEQ ID NO: 58LSLLFFVSGLFVNLVQAVCFVLIRPFSKNTYRRINRVVAELLWLELVWLIDWWAGVKIKVFTDHETLSLMGKEHALVISNHKSDIDWLVGWVLAQRSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYLFLERSWAKDESTLKSGLKRLKDYPLPFWLALFVEGTRFTQAKLLAAQQYAASSGLPVPRNVLIPRTKGFVSSVSHMRSFVPAIYDVTVAIPKMSTPPTMLRIFKGQSSVLHVHLKRHLMKDLPESDDAVAQWCRDIFVEKDALLDKHNAEDTFSGQELQDIGRPVKSLLVVISWAVLVIFGAVKFLQWSSLLSSWKGLAFSGIGLGIVTLLMHILILFSQSERSTPAKVAPAKPKKEGESSKTETEKEN CglutLPAAT1b SEQ ID NO: 59QAVCFVLIRPFSKNTYRRINRVVAELLWLELVWLIDWWAGVKIKVFTDHETLSLMGKEHALVISNHKSDIDWLVGWVLAQRSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYLFLERSWAKDESTLKSGLKRLKDYPLPFWLALFVEGTRFTQAKLLAAQQYAASSGLPVPRNVLIPRTKGFVSSVSHMRSFVPAIYDVTVAIPKMSTPPTMLRIFKGQSSVLHVHLKRHLMKDLPESDDAVAQWCRDIFVEKDALLDKHNAEDTFSGQELQDIGRPVKSLLVVISWAVLVIFGAVKFLQWSSLLSSWKGLAFSGIGLGIVTLLMHILILFSQSERSTPAKVAPAKPKKEGESSKTETEKEN CprLPAAT1SEQ ID NO: 60MAIAAAAVVFLFGLLFFTSGLIINLAQAVCFVLIWPLSKNAYRRINRVFAELLLLELLWLFHWRAGAKLKLFADPETFRLFGKEHALVICNHRTDLDWMVGWVLGQHFGCLGSILSVAKKSTKFLPVLGWSMWFSEYLFLERSWAKDKSTLKSHTERLKDYPLPFWLGIFVEGTRFTRAKLLAAQQYAASSGLPVPRNVLIPHTKGFVSSMSHMRSFVPAVYDLTVAFPKTSPPPTLLKLFEGQSVVLHVHIKRYAMKDLPESDDAVAQWCRDIYVEKDAFLDKHNAEDTFSGQEVHHIGRPIKSLLVVISWVVVIIFGALKFLRWSSLLSSWKGKAFSVIGLGIVTLLVNILILSSQAERSNPAKVVPAKLKTELSPSKKVTNKEN ChsLPAAT1 SEQ ID NO: 61MAIPSAAVVFLFGLLFFASGLIINLVQAVCFVLIWPLSKNTCRRINIVFQDMLLSELLWLFHWRAGAKLKFFTDPETYRHMGKEHALVITNHRTDLDWMIGWVLGEHLGCLGSILSVVKKSTKFLPVLGWSMWFSEYLFLERNWAKDKSTFKSHIERLEDFPQPFWFGIFVEGTRFTRAKLLAAQQYAASSGLPVPRNVLIPHTKGFVSSVSHMRSFVPAVYETTMTFPKTSPPPTLLKLFEGQPLVLHIHMKRHAMKDIPESDDAVAQWCRDKFVEKDALLDKHNAEDTFGGLEVHIGRSIKSLMVVICWVVVIIFGALKFLQWSSLLSSWKGIAFIGIGLGIVNLLVHVLILSSQAERSAPTKVAPAKLKTKLLSSKKITNKEN ChsLPAAT2 SEQ ID NO: 62MAIPSAAVVFLFGLLFFASGLIINLVQAVCFVLIWPLSKNTCRRINIVFQDMLLSELLWLFHWRAGAKLKFFTDPETYRHMGKEHALVITNHRTDLDWMIGWVLGEHLGCLGSILSVVKKSTKFLPVLGWSMWFSEYLFLERNWAKDKSTFKSHIERLEDFPQPFWFGIFVEGTRFTRAKLLAAQQYAASSGLPVPRNVLIPRTKGFVSSVSHMRSFVPAIYDVTVAIPKTSPPPTMLRMFKGQSSVLHVHLKRHLMKDLPESDDAVAQWCRDIFVEKDALLDKHNAEDTFSGQELQDIGRPIKSLVVVISWAALVVFGAVKFLQWSSLLSSWKGLAFSGIGLGIITLLMHILILFSQSERSTPAKVAPAKPKREGESSKTEMDKEN CcalcLPAAT1a SEQ ID NO: 63MAIPAAAVVFLFGLLFFPSGLIINLFQAVCFVLIWPFSRNTCRRINIVFQEMLLSELLWLFHWRAGAKLKLFADPETYRHMGKEHALLITNHRTDLDWMIGWALGQHLGCLGSILSVVKKSTKFLPSHIERLEDFPQPFWMAIFVEGTRFTRAKLLAAQQYAASSGLPVPRNVLIPRTKGFVSCVSHMRSFVPAVYETTMTFPKTSPPPTLLKLFEGQPIVLHVHMKRHAMKDIPESDEAVAQWCRDKFVEKDSLLDKHNAGDTFSCQEIHIGRPIKSLMVVISWVVVIIFGALKFLQWSSLLSSWKGIAFSGIGLGIVTLLVHILILSSQAERSTPAKVAPAKLKTELSSSTKVTNKENCcalcLPAAT1b SEQ ID NO: 64MAIPAAAVVFLFGLLFFPSGLIINLFQAVCFVLIWPFSRNTCRRINIVFQEMLLSELLWLFHWRAGAKLKLFADPETYRHMGKEHALLITNHRTDLDWMIGWALGQHLGCLGSILSVVKKSTKFLPVLGWSMWFSEYLFLERNWAKDKSTFKSHIERLEDFPQPFWMAIFVEGTRFTRAKLLAAQQYAASSGLPVPRNVLIPRTKGFVSCVSHMRSFVPAVYETTMTFPKTSPPPTLLKLFEGQPIVLHVHMKRHAMKDIPESDEAVAQWCRDKFVEKDSLLDKHNAGDTFSCQEIHIGRPIKSLMVVISWVVVIIFGALKFLQWSSLLSSWKGIAFSGIGLGIVTLLVHILILSSQAERSTPAKVAPAKLKTELSSSTKVTNKEN Cca1cLPAAT2 SEQ ID NO: 65LSLLFFVSGLIVNLVQAVCFVLIRPLSKNTYRRINRVVAELLWLELVWLIDWWAGVKIKVFTDHETFRLMGTEHALVISNHKSDIDWLVGWVLAQRSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYLFLERSWAKDESTLKSGLNRLKDYPLPFWLALFVEGTRFTRAKLLAAQQYAASSGLPVPRNVLIPRTKGFVSSVSHMRSFVPAIYDVTVAIPKTSPPPTMLRMFKGQSSVLHVHLKRHLMKDLPESDDAVAQWCRDIFVEKDALLDKHNAEDTFSGQELQDIGRPIKSLVVVISWAALVVFGAVKFLQWSSLLSSWKGLAFSGIALGIITLLMHILILFSQSERSTPAKVAPAKPKKEGESSKTETDKEN ChtLPAAT1a SEQ ID NO: 66MAIPAAAVIFLFSILFFASGLIINLVQAVCFVLIWPLSKNTCRRINLVFQEMLLSELLGLFHWRAGAKLKLYTDPETYPLLGKEHALLMINHRTDLDWMIGWVLGQHLGCLGSILSVVKKSTKFLPVLGWSMWFSEYLFLERNWAKDKSTFKSHIERLEDFPQPFWMAIFVEGTRFTRAKLLAAQQYAASSGLPVPRNVLIPHTKGFVSTVSHMRSFVPAVYDTTLTFPKTSPPPTLLNLFAGQPIVLHIHIKRHAMKDIPESDDAVAQWCRDKFVEKDALLDKHNAEDAFSDQEFPISRSIKSLMVVISWVMVIIFGALKFLQWSSLLSSWKGKAFSVIAVGIVTLLMHMSILSSQAERSNPAKVALPKLKTELPSSKKVLNKEN ChtLPAAT1b SEQ ID NO: 67MAIPAAAVIFLFSILFFASGLIINLVQAVCFVLIWPLSKNTCRRINLVFQEMLLSELLGLFHWRAGAKLKLYTDPETYPLLGKEHALLMINHRTDLDWMIGWVLGQHLGCLGSILSVVKKSTKFLPVLGWSMWFSEYLFLERNWAKDKSTFKSHIERLEDFPQPFWMAIFVEGTRFTRAKLLAAQQYAASSGLPVPRNVLIPHTKGFVSTVSHMRSFVPAVYDTTLTFPKTSPPPTLLNLFAGQPIVLHIHIKRHAMKDIPESDDAVAQWCRDKFVEKDALLDKHNAEDAFSDQEFPISRSIKSLMVVISWVMVIIFGALKFLQWSSLLSSWKGIAFSGIGLGIVTLLMHILILSSQAERSTPAKVAQAKVKTELPSSTKVTNKGN CwLPAAT1 SEQ ID NO: 68MAIPAAAVIFLFGILFFASGLIINLVQAVCFVLIWPLSKNTCRRINLVFQEMLLSELLWLFHWRAGAELKLFTDPETYRLLGKEHALVMTNHRTDLDWMIGWVTGQHLGCLGSILSIAKKSTKFLPVLGWSMWFSEYLFLERNWAKDKSTFKSHIERLEDFPQPFWMAIFVEGTRFTRAKLLAAQQYAASSGLPVPRNVLIPHTKGFVSSVCHMRSFVPAVYDTTLTFPKNSPPPTLLNLFAGQPIVLHIHIKRHAMKDMPKSDDAVAQWCRDKFVKKDALLDKHNTEDTFSDQEFPIGRPIKSLMVVISWVVVIIFGTLKFLQWSSLLSSWKGIAFSGIGLGIVTLLVHILILSSQAERSTPPKVAPAKLKTELSSTTKVINKGN CwLPAAT2b SEQ ID NO: 69LGLLFFVSGLIVNLVQAVCFVLIRPLSKNTYRRLNRVVAELLWLELVWLIDWWAGVKIKVFTDHETFHLMGKEHALVICNHKSDIDWLVGWVLAQRSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYLFLERSWAKDESTLKSGLNRLKDYPLPFWLALFVEGTRFTRAKLLAAQQYAASSGLPVPRNVLIPRTKGFVSSVSHMRSFVPAIYDVTVAIPKTSPPPTMLRMFKGQSSVDALLDKHNADDTFSGQELHDIGRPIKSLLVVISWAVLVVFGAVKFLQWSSLLSSWKGIAFSGIGLGIVTLLVHILILSSQAERSTSAKVAQAKVKTELSSSKKVKNKGN CwLPAAT2a SEQ ID NO: 70LGLLFFVSGLIVNLVQAVCFVLIRPLSKNTYRRLNRVVAELLWLELVWLIDWWAGVKIKVFTDHETFHLMGKEHALVICNHKSDIDWLVGWVLAQRSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYLFLERSWAKDESTLKSGLNRLKDYPLPFWLALFVEGTRFTRAKLLAAQQYAASSGLPVPRNVLIPRTKGFVSSVSHMRSFVPAIYDVTVAIPKTSPPPTMLRMFKGQSSVLHVHLKRHLMKDLPESDDAVAQWCRDIFVEKDVLLDKHNAEDTFSGQELQDIGRPVKSLLVVISWTLLVIFGAVKFLQWSSLLSSWKGLAFSGIGLGIVTLLMHILILFSQSERSTPAKVAPAKPKKEGESSKMETDKEN CgLPAAT1a SEQ ID NO: 71LAGWMGSSSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYLFLERSWAKDESTLKSGLNRLKDYPLPFWLALFVEGTRFTRAKLLAAQQYAASLGLPVPRNVLIPRTKGFVSSVSHMRSFVPAIYDVTVAIPKTSPPPTMIRMFKGQSSVLHVHLKRHVMKDLPESDDAVAQWCRDIFVEKDALLDKHNAEDTFSGQELQDTGRPIKSLLVVISWAVLEVFGAVKFLQWSSLLSSWKGLAFSGIGLGIITLLMHILILFSQSERSTPAKVAPAKPKNEGESSKAEMEKEKCgLPAAT1b SEQ ID NO: 72LAGWMGSSSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYLFLERSWAKDESTLKSGLNRLKDYPLPFWLALFVEGTRFTRAKLLAAQQYAASLGLPVPRNVLIPRTKGFVSSVSHMRSFVPAIYDVTVAIPKTSPPPTMIRMFKGQSSVLHVHLKRHVMKDLPESDDAVAQWCRDIFVEKDALLDKHNAEDTFSGQELQDTGRPIKSLLVRCFLVLSLIYLNGIMLKLRGPCLQVVISWAVLEVFGAVKFLQWSSLLSSWKGLAFSGIGLGIITLLMHILILFSQSERSTPAKVAPAKPKNEGESSKAEMEKEK CgLPAAT1c SEQ ID NO: 73LAGWMGSSSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYLFLERSWAKDESTLKSGLNRLKDYPLPFWLALFVEGTRFTRAKLLAAQQYAASLGLPVPRNVLIPRTKGFVSSVSHMRSFVPAIYDVTVAIPKTSPPPTMIRMFKGQSSVLHVHLKRHVMKDLPESDDAVAQWCRDIFVEKDALLDKHNAEDTFSGQELQDTGRPIKSLLVVTSWAVLVISGAVKFLQWSSLLSSWKGLAFSGIGLGIVTLLMHILILFSQSERSTPAKVAPAKPKKEGESSKTEKDKENCpalLPAAT1 SEQ ID NO: 74LGLLFFVSGLIVNLVQAVCFVLIRPLSKNTYRRINRVVAELLWLELVWLIDWWAGVKIKVFTDHETLSLMGKEHALVICNHKSDIDWLVGWVLAQRSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYLFLERSWAKDENTLKSGLNRLKDYPLPFWLALFVEGTRFTRAKLLAAQQYATSSGLPVPRNVLIPRTKGFVSSVSHMRSFVPAIYDVTVAIPKTSPPPTMLRMFKGQSSVLHVHLKRHLMKDLPESDDAVAQWCRDIFVEKDALLDKHNAEDTFSGQELQDTGRPIKSLLVVISWAVLVIFGAVKFLQWSSLLSSWKGLAFSGVGLGIITLLMHILILFSQSERSTPAKVAPAKPKKDGESSKTEIEKEN CaLPAAT1 SEQ ID NO: 75MAIAAAAVIVPVSLLFFVSGLIVNLVQAVCFVLIRPLFKNTYRRINRVVAELLWLELVWLIDWWAGVKIKVFTDHETFHLMGKEHALVICNHKSDIDWLVGWVLAQRSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYLFLERNWAKDESTLKSGLNRLKDYPLPFWLALFVEGTRFTRAKLLAAQQYAASSGLPVPRNVLIPRTKGFVSSVSHMRSFVPAIYDVTVAIPKTSPPPTLLRMFKGQSSVLHVHLKRHQMNDLPESDDAVAQWCRDIFVEKDALLDKHNAEDTFSGQELQDTGRPIKSLLIVISWAVLVVFGAVKFLQWSSLLSSWKGLAFSGIGLGVITLLMHILILFSQSERSTPAKVAPAKPKIEGESSKTEMEKEH CaLPAAT3 SEQ ID NO: 76MTIASAAVVFLFGILLFTSGLIINLFQAFCSVLVWPLSKNAYRRINRVFAEFLPLEFLWLFHWWAGAKLKLFTDPETFRLMGKEHALVIINHKIELDWMVGWVLGQHLGCLGSILSVAKKSTKFLPVFGWSLWFSEYLFLERNWAKDKKTLKSHIERLKDYPLPFWLIIFVEGTRFTRTKLLAAQQYAASAGLPVPRNVLIPHTKGFVSSVSHMRSFVPAIYDVTVAFPKTSPPPTMLKLFEGHFVELHVHIKRHAMKDLPESEDAVAQWCRDKEVEKDALLDKHNAEDTFSGQEVHHVGRPIKSLLVVISWVVVIIFGALKFLQWSSLLSSWKGIAFSVIGLGTVALLMQILILSSQAERSIPAKETPANLKTELSSSKKVTNKEN SalL2AAT1 SEQ ID NO: 77MAIGAAAIVVPLGLLFMLSGLMVNLIQAICFILVRPLSKNMYRRVNRVVVELLWLELIWLIDWWGGVKVDVYADSETFQSLGKEHALVVSNHKSDIDWLVGWVLAQRSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYVFLERSWAKDESTLKSGLQRLKDFPRPFWLALFVEGTRFTQAKLLAAQEYAASTGLPIPRNVLIPRTKGFVSAVSNMRSFVPAIYDVTVAIPKTQPSPTMLRIFNRQPSVVHVRIKRHSMNQLPPTDEGVAQWCKDIFVAKDALLDRHLAEGKFDEKEFKRIRRPIKSLLVISSWSFLLLFGVFKFLKWSALLSTWKGVAVSTAVLLLVTVVMYMFILFSQSERSSPRKVAPSGPENGC1eptL2AAT1 SEQ ID NO: 78MAIPAAVVIFLFGLLFFSSGLIINLFQALCFVLIWPLSKNAYRRINRVFAELLLSELLCLFDWWAGAKLKLFTDPETFRLMGKEHALVIINHMTELDWMVGWVMGQHFGCLGSILSVAKKSTKFLPVLGWSMWFTEYLYIERSWDKDKSTLKSHIERLKDYPLPFWLVIFAEGTRFTRTKLLAAQQYAASSGLPVPRNVLIPRTKGFVSCVNHMRSFVPAVYDLTVAFPKTSPPPTLLNLFEGQSVVLHVHIKRHAMKDLPESDDAVAQWCRDKFVEKDALLDKHNAEDTFSSQEVHHTGSRPIKSLLVVISWVVVITFGALKFLQWSSWKGKAFSVIGLGIVTLLMHMLILSSQAERSKPAKVTQAKLKTELSISKKVTDKEN ClopLPAAT1 SEQ ID NO: 79MAIAAAAVIFLFGLLFFASGLIINLFQALCFVLIRPLSKNAYRRINRVFAELLLSELLCLFDWWAGAKLKLFTDPETLRLMGKEHALIIINHMTELDWMVGWVMGQHFGCLGSIISVAKKSTKFLPVLGWSMWFSEYLYLERSWAKDKSTLKSHIERLKDYPLPFWLVIFVEGTRFTRTKLLAAQEYAASSGLPVPRNVLIPRTKGFVSCVNHMRSFVPAVYDVTVAFPKTSPQPTLLNLFEGRSIVLHVHIKRHAMKDLPESDDAVAQWCRDKFVEKDALLDKHNAEDTFSGQEVHHTGRRPIKSLLVVMSWVVVTTFGALKFLQWSSWKGKAFSVIGLGIVTLLMHVLILSSQAERSNPAKVVQAELNTELSISKKVTNKGN CcrasLPAAT1a SEQ ID NO: 80MAIPAAAVIFLFGLIFFASGLIINLFQALCFVLIWPLWKNAYRRINRVFAELLLSELLCLFDWWAGAKLKLFTDPETFRLMGKEHALVIINHMTELDWMVGWVMGQHFGCLGSILSVAKKSTKFLPVLGWSMWFTEYLYIERSWDKDKSTLKSHIERLKDYPLPFWLVIFAEGTRFTRTKLLAAQQYAASSGLPVPRNVLIPRTKGFVSSVSHMRSFVPAIYDVTVAIPKTSPPPTLIRMFKGQSSVLHVHLKRHVMKDLPESDDAVAQWCRDIFVEKDALLDKHNAEDTFSGQELQDTGRPIKSLLVVISWAVLEVFGAVKFLQWSSLLSSWKGLAFSGIGLGIITLLMHILILFSQSERSTPAKVAPAKAKCcrasLPAAT1b SEQ ID NO: 81MAIPAAAVIFLFGLIFFASGLIINLFQALCFVLIWPLWKNAYRRINRVFAELLLSELLCLFDWWAGAKLKLFTDPETFRLMGKEHALVIINHMTELDWMVGWVMGQHFGCLGSILSVAKKSTKFLPVLGWSMWFTEYLYIERSWDKDKSTLKSHIERLKDYPLPFWLVIFAEGTRFTRTKLLAAQQYAASSGLPVPRNVLIPRTKGFVSSVSHMRSFVPAIYDVTVAIPKTSPPPTLIRMFKGQSSVLHVHLKRHVMKDLPESDDAVAQWCRDIFVEKDALLDKHNAEDTFSGQELQDTGRPIKSLLVRCFLVLSLIYLNGIILKLCGLCLQVVISWAVLEVFGAVKFLQWSSLLSSWKGLAFSGIGLGIITLLMHILILFSQSERSTPAKVAPAKAKSEQ ID NO: CcrasLPAAT1c SEQ ID NO: 82MAIPAAAVIFLFGLIFFASGLIINLFQALCFVLIWPLWKNAYRRINRVFAELLLSELLCLFDWWAGAKLKLFTDPETFRLMGKEHALVIINHMTELDWMVGWVMGQHFGCLGSILSVAKKSTKFLPVLGWSMWFTEYLYIERSWDKDKSTLKSHIERLKDYPLPFWLVIFAEGTRFTRTKLLAAQQYAASSGLPVPRNVLIPRTKGFVSSVSHMRSFVPAIYDVTVAIPKTSPPPTLIRMFKGQSSVLHVHLKRHVMKDLPESDDAVAQWCRDIFVEKDALLDKHNAEDTFSGQELQDTGRPIKSLLVVISWAVLEVFGAVKFLQWSSLLSSWKGLAFSGIGLGIITLLMHILILFSQSERSTPAKVAPAKAKMEGESSKTEMEMEK CcrasLPAAT1d SEQ ID NO: 83MAIPAAAVIFLFGLIFFASGLIINLFQALCFVLIWPLWKNAYRRINRVFAELLLSELLCLFDWWAGAKLKLFTDPETFRLMGKEHALVIINHMTELDWMVGWVMGQHFGCLGSILSVAKKSTKFLPVLGWSMWFTEYLYIERSWDKDKSTLKSHIERLKDYPLPFWLVIFAEGTRFTRTKLLAAQQYAASSGLPVPRNVLIPRTKGFVSSVSHMRSFVPAIYDVTVAIPKTSPPPTLIRMFKGQSSVLHVHLKRHVMKDLPESDDAVAQWCRDIFVEKDALLDKHNAEDTFSGQELQDTGRPIKSLLVRCFLVLSLIYLNGIILKLCGLCLQVVISWAVLEVFGAVKFLQWSSLLSSWKGLAFSGIGLGIITLLMHILILFSQSERSTPAKVAPAKAKMEGESSKTEMEMEK CkoeLPAAT1 SEQ ID NO: 84MAIAAAPVIFLFGLLFFASGLIINLFQAICFVLIWPLSKNAYRRINRVFAELLLSELLCLFDWWAGAKLKLFTDPETFRLMGKEHALVITNHKIDLDWMIGWILGQHFGCLGSVISIAKKSTKFLPIFGWSLWFSEYLFLERNWAKDKRTLKSHIERMKDYPLPLWLILFVEGTRFTRTKLLAAQQYAASSGLPVPRNVLIPHTKGFVSSVSHMRSFVPAIYDVTVAIPKTSPPPTLIRMFKGQSSVLHVHLKRHLMKDLPESDDAVAQWCRDIFVEKDALLDKHNAEDTFSGQELQETGRPIKSLLVVISWAVLEVYGAVKFLQWSSLLSSWKGLAFSGIGLGLITLLMHILILFSQSERSTPAKVAPAKPKKEGESSKTEMEKEK CkoeLPAAT2 SEQ ID NO: 85MHVLLEMVTFRFSSFFVFDNVQALCFVLIWPLSKSAYRKINRVFAELLLSELLCLFDWWAGAKLKLFTDPETFRLMGKEHALVITNHKIDLDWMIGWILGQHFGCLGSVISIAKKSTKFLPIFGWSLWFSEYLFLERNWAKDKRTLKSHIERMKDYPLPLWLILFVEGTRFTRTKLLAAQQYAASSGLPVPRNVLIPHTKGFVSSVSHMRSFVPAVYDVTVAFPKTSPPPTMLSLFEGQSVVLHVHIKRHAMKDLPDSDDAVAQWCRDKFVEKDALLDKHNAEDTFSGQEVHHVGRPIKSLLVVISWMVVIIFGALKFLQWSSLLSSWKGKAFSAIGLGIATLLMHVLVVFSQADRSNPAKVPPAKLNTELSSSKKVTNKEN pSZ2624 PmKASII SEQ ID NO: 86 gtttaaacGCCGGTCACCACCCGCATGCTCGTACTACAGCGCACGCACCGCTTCGTGATCCACCGGGTGAACGTAGTCCTCGACGGAAACATCTGGTTCGGGCCTCCTGCTTGCACTCCCGCCCATGCCGACAACCTTTCTGCTGTTACCACGACCCACAATGCAACGCGACACGACCGTGTGGGACTGATCGGTTCACTGCACCTGCATGCAATTGTCACAAGCGCTTACTCCAATTGTATTCGTTTGTTTTCTGGGAGCAGTTGCTCGACCGCCCGCGTCCCGCAGGCAGCGATGACGTGTGCGTGGCCTGGGTGTTTCGTCGAAAGGCCAGCAACCCTAAATCGCAGGCGATCCGGAGATTGGGATCTGATCCGAGTTTGGACCAGATCCGCCCCGATGCGGCACGGGAACTGCATCGACTCGGCGCGGAACCCAGCTTTCGTAAATGCCAGATTGGTGTCCGATACCTGGATTTGCCATCAGCGAAACAAGACTTCAGCAGCGAGCGTATTTGGCGGGCGTGCTACCAGGGTTGCATACATTGCCCATTTCTGTCTGGACCGCTTTACTGGCGCAGAGGGTGAGTTGATGGGGTTGGCAGGCATCGAAACGCGCGTGCATGGTGTGCGTGTCTGTTTTCGGCTGCACGAATTCAATAGTCGGATGGGCGACGGTAGAATTGGGTGTGGCGCTCGCGTGCATGCCTCGCCCCGTCGGGTGTCATGACCGGGACTGGAATCCCCCCTCGCGACCATCTTGCTAACGCTCCCGACTCTCCCGACCGCGCGCAGGATAGACTCTTGTTCAACCAATCGACA actagt ATGcagaccgcccaccagcgcccccccaccgagggccactgcttcggcgcccgcctgcccaccgcctcccgccgcgccgtgcgccgcgcctggtcccgcatcgcccgcg ggcgcgccgccgccgccgccgacgccaaccccgcccgccccgagcgccgcgtggtgatcaccggccagggcgtggtgacctccctgggccagaccatcgagcagactactcctccctgctggagggcgtgtccggcatctcccagatccagaagacgacaccaccggctacaccaccaccatcgccggcgagatcaagtccctgcagctggacccctacgtgcccaagcgctgggccaagcgcgtggacgacgtgatcaagtacgtgtacatcgccggcaagcaggccctggagtccgccggcctgcccatcgaggccgccggcctggccggcgccggcctggaccccgccctgtgcggcgtgctgatcggcaccgccatggccggcatgacctccacgccgccggcgtggaggccctgacccgcggcggcgtgcgcaagatgaaccccactgcatccccactccatctccaacatgggcggcgccatgctggccatggacatcggatcatgggccccaactactccatctccaccgcctgcgccaccggcaactactgcatcctgggcgccgccgaccacatccgccgcggcgacgccaacgtgatgctggccggcggcgccgacgccgccatcatcccctccggcatcggcggcttcatcgcctgcaaggccctgtccaagcgcaacgacgagcccgagcgcgcctcccgcccctgggacgccgaccgcgacggatcgtgatgggcgagggcgccggcgtgctggtgctggaggagctggagcacgccaagcgccgcggcgccaccatcctggccgagctggtgggcggcgccgccacctccgacgcccaccacatgaccgagcccgacccccagggccgcggcgtgcgcctgtgcctggagcgcgccctggagcgcgcccgcctggcccccgagcgcgtgggctacgtgaacgcccacggcacctccacccccgccggcgacgtggccgagtaccgcgccatccgcgccgtgatcccccaggactccctgcgcatcaactccaccaagtccatgatcggccacctgctgggcggcgccggcgccgtggaggccgtggccgccatccaggccctgcgcaccggctggctgcaccccaacctgaacctggagaaccccgcccccggcgtggaccccgtggtgctggtgggcccccgcaaggagcgcgccgaggacctggacgtggtgctgtccaactccttcggcttcggcggccacaactcctgcgtgatcttccgcaagtacgacgagatggactacaag gaccacgacggcgactacaaggaccacgacatcgactacaaggatcgacgacgacaagTGA atcgatAGATCTCTTAAGGCAGCAGCAGCTCGGATAGTATCGACACACTCTGGACGCTGGTCGTGTGATGGACTGTTGCCGCCACACTTGCTGCCTTGACCTGTGAATATCCCTGCCGCTTTTATCAAACAGCCTCAGTGTGTTTGATCTTGTGTGTACGCGCTTTTGCGAGTTGCTAGCTGCTTGTGCTATTTGCGAATACCACCCCCAGCATCCCCTTCCCTCGTTTCATATCGCTTGCATCCCAACCGCAACTTATCTACGCTGTCCTGCTATCCCTCAGCGCTGCTCCTGCTCCTGCTCACTGCCCCTCGCACAGCCTTGGTTTGGGCTCCGCCTGTATTCTCCTGGTACTGCAACCTGTAAACCAGCACTGCAATGCTGATGCACGGGAAGTAGTGGGATGGGAACACAAATGGAAAGCTTAATTAAgagctccgcgtctcgaacagagcgcgcagaggaacgctgaaggtctcgcctctgtcgcacctcagcgcggcatacaccacaataaccacctgacgaatgcgcttggttcttcgtccattagcgaagcgtccggttcacacacgtgccacgttggcgaggtggcaggtgacaatgatcggtggagctgatggtcgaaacgttcacagcctaggtgatatccatcttaaggatctaagtaagattcgaagcgctcgaccgtgccggacggactgcagccccatgtcgtagtgaccgccaatgtaagtgggctggcgtaccctgtacgtgagtcaacgtcactgcacgcgcaccaccctctcgaccggcaggaccaggcatcgcgagatacagcgcgagccagacacggagtgccgagctatgcgcacgctccaactaggtaccagtttaggtccagcgtccgtggggggggacgggctgggagcttgggccgggaagggcaagacgatgcagtccctctggggagtcacagccgactgtgtgtgttgcactgtgcggcccgcagcactcacacgcaaaatgcctggccgacaggcaggccctgtccagtgcaacatccacggtccctctcatcaggctcaccttgctcattgacataacggaatgcgtaccgctctttcagatctgtccatccagagaggggagcaggctccccaccgacgctgtcaaacttgcttcctgcccaaccgaaaacattattgtttgagggggggggggggggggcagattgcatggcgggatatctcgtgaggaacatcactgggacactgtggaacacagtgagtgcagtatgcagagcatgtatgctaggggtcagcgcaggaagggggcctttcccagtctcccatgccactgcaccgtatccacgactcaccaggaccagcttcttgatcggcttccgctcccgtggacaccagtgtgtagcctctggactccaggtatgcgtgcaccgcaaaggccagccgatcgtgccgattcctgggtggaggatatgagtcagccaacttggggctcagagtgcacactggggcacgatacgaaacaacatctacaccgtgtcctccatgctgacacaccacagcttcgctccacctgaatgtgggcgcatgggcccgaatcacagccaatgtcgctgctgccataatgtgatccagaccctctccgcccagatgccgagcggatcgtgggcgctgaatagattcctgtttcgatcactgtttgggtcctttccillicgtctcggatgcgcgtctcgaaacaggctgcgtcgggctttcggatcccttttgctccctccgtcaccatcctgcgcgcgggcaagttgcttgaccctgggctgataccagggttggagggtattaccgcgtcaggccattcccagcccggattcaattcaaagtctgggccaccaccctccgccgctctgtctgatcactccacattcgtgcatacactacgttcaagtcctgatccaggcgtgtctcgggacaaggtgtgcttgagtttgaatctcaaggacccactccagcacagctgctggttgaccccgccctcgcaatctagaATGgccgcgtccgtccactgcaccctgatgtccgtggtctgcaacaacaagaaccactccgcccgccccaagctgcccaactcctccctgctgcccggatcgacgtggtggtccaggccgcggccacccgatcaagaaggagacgacgaccacccgcgccacgctgacgacgacccccccacgaccaactccgagcgcgccaagcagcgcaagcacaccatcgacccctcctcccccgacaccagcccatcccctccacgaggagtgatccccaagtccacgaaggagcacaaggaggtggtgcacgaggagtccggccacgtcctgaaggtgcccaccgccgcgtgcacctgtccggcggcgagcccgccacgacaactacgacacgtccggcccccagaacgtcaacgcccacatcggcctggcgaagctgcgcaaggagtggatcgaccgccgcgagaagctgggcacgccccgctacacgcagatgtactacgcgaagcagggcatcatcacggaggagatgctgtactgcgcgacgcgcgagaagctggaccccgagacgtccgctccgaggtcgcgcggggccgcgccatcatcccctccaacaagaagcacctggagctggagcccatgatcgtgggccgcaagacctggtgaaggtgaacgcgaacatcggcaactccgccgtggcctcctccatcgaggaggaggtctacaaggtgcagtgggccaccatgtggggcgccgacaccatcatggacctgtccacgggccgccacatccacgagacgcgcgagtggatcctgcgcaactccgcggtccccgtgggcaccgtccccatctaccaggcgctggagaaggtggacggcatcgcggagaacctgaactgggaggtgaccgcgagacgctgatcgagcaggccgagcagggcgtggactacttcacgatccacgcgggcgtgctgctgcgctacatccccctgaccgccaagcgcctgacgggcatcgtgtcccgcggcggctccatccacgcgaagtggtgcctggcctaccacaaggagaacttcgcctacgagcactgggacgacatcctggacatctgcaaccagtacgacgtcgccctgtccatcggcgacggcctgcgccccggctccatctacgacgccaacgacacggcccagacgccgagctgctgacccagggcgagctgacgcgccgcgcgtgggagaaggacgtgcaggtgatgaacgagggccccggccacgtgcccatgcacaagatccccgagaacatgcagaagcagctggagtggtgcaacgaggcgcccactacaccctgggccccctgacgaccgacatcgcgcccggctacgaccacatcacctccgccatcggcgcggccaacatcggcgccctgggcaccgccctgctgtgctacgtgacgcccaaggagcacctgggcctgcccaaccgcgacgacgtgaaggcgggcgtcatcgcctacaagatcgccgcccacgcggccgacctggccaagcagcacccccacgcccaggcgtgggacgacgcgctgtccaaggcgcgatcgagaccgctggatggaccagacgcgctgtccctggaccccatgacggcgatgtccaccacgacgagacgctgcccgcggacggcgcgaaggtcgcccacactgctccatgtgcggccccaagactgctccatgaagatcacggaggacatccgcaagtacgccgaggagaacggctacggctccgccgaggaggccatccgccagggcatggacgccatgtccgaggagacaacatcgccaagaagacgatctccggcgagcagcacggcgaggtcggcggcgagatctacctgcccgagtcctacgtcaaggccgcgcagaagTGA caattgACGGAGCGTCGTGCGGGAGGGAGTGTGCCGAGCGGGGAGTCCCGGTCTGTGCGAGGCCCGGCAGCTGACGCTGGCGAGCCGTACGCCCCGAGGGTCCCCCTCCCCTGCACCCTCTTCCCCTTCCCTCTGACGGCCGCGCCTGTTCTTGCATGTTCAGCGACggatcc TAGGGAGCGACGAGTGTGCGTGCGGGGCTGGCGGGAGTGGGACGCCCTCCTCGCTCCTCTCTGTTCTGAACGGAACAATCGGCCACCCCGCGCTACGCGCCACGCATCGAGCAACGAAGAAAACCCCCCGATGATAGGTTGCGGTGGCTGCCGGGATATAGATCCGGCCGCACATCAAAGGGCCCCTCCGCCAGAGAAGAAGCTCCTTTCCCAGCAGACTCCTTCTGCTGCCAAAACACTTCTCTGTCCACAGCAACACCAAAGGATGAACAGATCAACTTGCGTCTCCGCGTAGCTTCCTCGGCTAGCGTGCTTGCAACAGGTCCCTGCACTATTATCTTCCTGCTTTCCTCTGAATTATGCGGCAGGCGAGCGCTCGCTCTGGCGAGCGCTCCTTCGCGCCGCCCTCGCTGATCGAGTGTACAGTCAATGAATGGTCCTGGGCGAAGAACGAGGGAATTTGTGGGTAAAACAAGCATCGTCTCTCAGGCCCCGGCGCAGTGGCCGTTAAAGTCCAAGACCGTGACCAGGCAGCGCAGCGCGTCCGTGTGCGGGCCCTGCCTGGCGGCTCGGCGTGCCAGGCTCGAGAGCAGCTCCCTCAGGTCGCCTTGGACGGCCTCTGCGAGGCCGGTGAGGGCCTGCAGGAGCGCCTCGAGCGTGGCAGTGGCGGTCGTATCCGGGTCGCCGGTCACCGCCTGCGACTCGCCATCCgaagagcgtttaaac pSZ3204 GarmFATA SEQ ID NO: 87gctettc GCCGCCGCCACTCCTGCTCGAGCGCGCCCGCGCGTGCGCCGCCAGCGCCTTGGCCTTTTCGCCGCGCTCGTGCGCGTCGCTGATGTCCATCACCAGGTCCATGAGGTCTGCCTTGCGCCGGCTGAGCCACTGCTTCGTCCGGGCGGCCAAGAGGAGCATGAGGGAGGACTCCTGGTCCAGGGTCCTGACGTGGTCGCGGCTCTGGGAGCGGGCCAGCATCATCTGGCTCTGCCGCACCGAGGCCGCCTCCAACTGGTCCTCCAGCAGCCGCAGTCGCCGCCGACCCTGGCAGAGGAAGACAGGTGAGGGGGGTATGAATTGTACAGAACAACCACGAGCCTTGTCTAGGCAGAATCCCTACCAGTCATGGCTTTACCTGGATGACGGCCTGCGAACAGCTGTCCAGCGACCCTCGCTGCCGCCGCTTCTCCCGCACGCTTCTTTCCAGCACCGTGATGGCGCGAGCCAGCGCCGCACGCTGGCGCTGCGCTTCGCCGATCTGAGGACAGTCGGGGAACTCTGATCAGTCTAAACCCCCTTGCGCGTTAGTGTTGCCATCCTTTGCAGACCGGTGAGAGCCGACTTGTTGTGCGCCACCCCCCACACCACCTCCTCCCAGACCAATTCTGTCACCTTTTTGGCGAAGGCATCGGCCTCGGCCTGCAGAGAGGACAGCAGTGCCCAGCCGCTGGGGGTTGGCGGATGCA CGCTCAggtaccctttcttgcgctatgacacttccagcaaaaggtagggcgggctgcgagacggcttcccggcgctgcatgcaacaccgatgatgcttcgaccccccgaagctccttcggggctgcatgggcgctccgatgccgctccagggcgagcgctgtttaaatagccaggcccccgattgcaaagacattatagcgagctaccaaagccatattcaaacacctagatcactaccacttctacacaggccactcgagcttgtgatcgcactccgctaagggggcgcctcttcctcttcgtttcagtcacaacccgcaaactctagaatatcaATGctgctgcaggccttcctgttcctgctggcggcttcgccgccaagatcagcgcctccatgacgaacgagacgtccgaccgccccctggtgcacttcacccccaacaagggctggatgaacgaccccaacggcctgtggtacgacgagaaggacgccaagtggcacctgtacaccagtacaacccgaacgacaccgtctgggggacgccatgactggggccacgccacgtccgacgacctgaccaactgggaggaccagcccatcgccatcgccccgaagcgcaacgactccggcgccactccggctccatggtggtggactacaacaacacctccggcacttcaacgacaccatcgacccgcgccagcgctgcgtggccatctggacctacaacaccccggagtccgaggagcagtacatctcctacagcctggacggcggctacaccacaccgagtaccagaagaaccccgtgctggccgccaactccacccagaccgcgacccgaaggtcactggtacgagccctcccagaagtggatcatgaccgcggccaagtcccaggactacaagatcgagatctactcctccgacgacctgaagtcctggaagctggagtccgcgacgccaacgagggcacctcggctaccagtacgagtgccccggcctgatcgaggtccccaccgagcaggaccccagcaagtcctactgggtgatgacatctccatcaaccccggcgccccggccggcggctcatcaaccagtacttcgtcggcagatcaacggcacccacttcgaggccacgacaaccagtcccgcgtggtggacttcggcaaggactactacgccctgcagaccacttcaacaccgacccgacctacgggagcgccctgggcatcgcgtgggcctccaactgggagtactccgccacgtgcccaccaacccctggcgctcctccatgtccctcgtgcgcaagactccctcaacaccgagtaccaggccaacccggagacggagctgatcaacctgaaggccgagccgatcctgaacatcagcaacgccggcccctggagccggacgccaccaacaccacgagacgaaggccaacagctacaacgtcgacctgtccaacagcaccggcaccctggagacgagctggtgtacgccgtcaacaccacccagacgatctccaagtccgtgacgcggacctctccctctggacaagggcctggaggaccccgaggagtacctccgcatgggatcgaggtgtccgcgtcctccacacctggaccgcgggaacagcaaggtgaagacgtgaaggagaacccctacttcaccaaccgcatgagcgtgaacaaccagccatcaagagcgagaacgacctgtcctactacaaggtgtacggcagctggaccagaacatcctggagctgtacttcaacgacggcgacgtcgtgtccaccaacacctacttcatgaccaccgggaacgccctgggctccgtgaacatgacgacgggggtggacaacctgttctacatcgacaagttccaggtgcgcgaggtcaagTGAcaattgGCAGCA GCAGCTCGGATAGTATCGACACACTCTGGACGCTGGTCGTGTGATGGACTGTTGCCGCCACACTTGCTGCCTTGACCTGTGAATATCCCTGCCGCTTTTATCAAACAGCCTCAGTGTGTTTGATCTTGTGTGTACGCGCTTTTGCGAGTTGCTAGCTGCTTGTGCTATTTGCGAATACCACCCCCAGCATCCCCTTCCCTCGTTTCATATCGCTTGCATCCCAACCGCAACTTATCTACGCTGTCCTGCTATCCCTCAGCGCTGCTCCTGCTCCTGCTCACTGCCCCTCGCACAGCCTTGGTTTGGGCTCCGCCTGTATTCTCCTGGTACTGCAACCTGTAAACCAGCACTGCAATGCTGATGCACGGGAAGTAGTGGGATGGGAACACAAATGGAggatcccgcgtctcgaacagagcgcgcagaggaacgctgaaggtctcgcctctgtcgcacctcagcgcggcatacaccacaataaccacctgacgaatgcgcttggttcttcgtccattagcgaagcgtccggttcacacacgtgccacgttggcgaggtggcaggtgacaatgatcggtggagctgatggtcgaaacgttcacagcctagggatatcctgaagaatgggaggcaggtgttgttgattatgagtgtgtaaaagaaaggggtagagagccgtcctcagatccgactactatgcaggtagccgctcgcccatgcccgcctggctgaatattgatgcatgcccatcaaggcaggcaggcatactgtgcacgcaccaagcccacaatcttccacaacacacagcatgtaccaacgcacgcgtaaaagttggggtgctgccagtgcgtcatgccaggcatgatgtgctcctgcacatccgccatgatctcctccatcgtctcgggtgtttccggcgcctggtccgggagccgttccgccagatacccagacgccacctccgacctcacggggtactittcgagcgtctgccggtagtcgacgatcgcgtccaccatggagtagccgaggcgccggaactggcgtgacggagggaggagagggaggagagagaggggggggggggggggggatgattacacgccagtctcacaacgcatgcaagacccgtttgattatgagtacaatcatgcactactagatggatgagcgccaggcataaggcacaccgacgttgatggcatgagcaactcccgcatcatatttcctattgtcctcacgccaagccggtcaccatccgcatgctcatattacagcgcacgcaccgcttcgtgatccaccgggtgaacgtagtcctcgacggaaacatctggctcgggcctcgtgctggcactccctcccatgccgacaacctttctgctgtcaccacgacccacgatgcaacgcgacacgacccggtgggactgatcggttcactgcacctgcatgcaattgtcacaagcgcatactccaatcgtatccgtttgatttctgtgaaaactcgctcgaccgcccgcgtcccgcaggcagcgatgacgtgtgcgtgacctgggtgtttcgtcgaaaggccagcaaccccaaatcgcaggcgatccggagattgggatctgatccgagcttggaccagatcccccagatgcggcacgggaactgcatcgactcggcgcggaacccagctttcgtaatgccagattggtgtccgataccttgatttgccatcagcgaaacaagacttcagcagcgagcgtatttggcgggcgtgctaccagggttgcatacattgcccatttctgtctggaccgctttaccggcgcagagggtgagttgatggggttggcaggcatcgaaacgcgcgtgcatggtgtglgtgtctgttttcggctgcacaatttcaatagtcggatgggcgacggtagaattgggtgttgcgctcgcgtgcatgcctcgccccgtcgggtgtcatgaccgggactggaatcccccctcgcgaccctcctgctaacgctcccgactctcccgcccgcgcgcaggatagactctagttcaaccaatcgacaactagt ATGgccaccgcatccactactcggcgacaatgcccgctgcggcgacctgcgtcgctcggcgggctccgggccccggcgcccagcgaggcccctccccgtgcgc gggcgcgccatccccccccgcatcatcgtggtgtcctcctcctcctccaaggtgaaccccctgaagaccgaggccgtggtgtcctccggcctggccgaccgcctgcgcctgggctccctgaccgaggacggcctgtcctacaaggagaagacatcgtgcgctgctacgaggtgggcatcaacaagaccgccaccgtggagaccatcgccaacctgctgcaggaggtgggctgcaaccacgcccagtccgtgggctactccaccggcggcactccaccacccccaccatgcgcaagctgcgcctgatctgggtgaccgcccgcatgcacatcgagatctacaagtaccccgcctggtccgacgtggtggagatcgagtcctggggccagggcgagggcaagatcggcacccgccgcgactggatcctgcgcgactacgccaccggccaggtgatcggccgcgccacctccaagtgggtgatgatgaaccaggacacccgccgcctgcagaaggtggacgtggacgtgcgcgacgagtacctggtgcactgcccccgcgagctgcgcctggccaccccgaggagaacaactcctccctgaagaagatctccaagctggaggacccctcccagtactccaagctgggcctggtgccccgccgcgccgacctggacatgaaccagcacgtgaacaacgtgacctacatcggctgggtgctggagtccatgccccaggagatcatcgacacccacgagctgcagaccatcaccctggactaccgccgcgagtgccagcacgacgacgtggtggactccctgacctcccccgagccctccgaggacgccgaggccgtgacaaccacaacggcaccaacggctccgccaacgtgtccgccaacgaccacggctgccgcaacacctgcacctgctgcgcctgtccggcaacggcctggagatcaaccgcggccgcaccgagtggcgcaagaagcccacccgcATGGACTACAAGGACCACGACGGCGACTACAAGGACCACGACATCGACTACAAGGACGACGACGACAAGTGA ategatagatctcttaagGCAGCAGCAGCTCGGATAGTATCGACACACTCTGGACGCTGGTCGTGTGATGGACTGTTGCCGCCACACTTGCTGCCTTGACCTGTGAATATCCCTGCCGCTTTTATCAAACAGCCTCAGTGTGTTTGATCTTGTGTGTACGCGCTTTTGCGAGTTGCTAGCTGCTTGTGCTATTTGCGAATACCACCCCCAGCATCCCCTTCCCTCGTTTCATATCGCTTGCATCCCAACCGCAACTTATCTACGCTGTCCTGCTATCCCTCAGCGCTGCTCCTGCTCCTGCTCACTGCCCCTCGCACAGCCTTGGTTTGGGCTCCGCCTGTATTCTCCTGGTACTGCAACCTGTAAACCAGCACTGCAATGCTGATGCACGGGAAGTAGTGGGATGGGAACACAAATGGAaagcttaattaagagctc TTGTTTTCCAGAAGGAGTTGCTCCTTGAGCCTTTCATTCTCAGCCTCGATAACCTCCAAAGCCGCTCTAATTGTGGAGGGGGTTCGAATTTAAAAGCTTGGAATGTTGGTTCGTGCGTCTGGAACAAGCCCAGACTTGTTGCTCACTGGGAAAAGGACCATCAGCTCCAAAAAACTTGCCGCTCAAACCGCGTACCTCTGCTTTCGCGCAATCTGCCCTGTTGAAATCGCCACCACATTCATATTGTGACGCTTGAGCAGTCTGTAATTGCCTCAGAATGTGGAATCATCTGCCCCCTGTGCGAGCCCATGCCAGGCATGTCGCGGGCGAGGACACCCGCCACTCGTACAGCAGACCATTATGCTACCTCACAATAGTTCATAACAGTGACCATATTTCTCGAAGCTCCCCAACGAGCACCTCCATGCTCTGAGTGGCCACCCCCCGGCCCTGGTGCTTGCGGAGGGCAGGTCAACCGGCATGGGGCTACCGAAATCCCCGACCGGATCCCACCACCCCCGCGATGGGAAGAATCTCTCCCCGGGATGTGGGCCCACCACCAGCACAACCTGCTGGCCCAGGCGAGCGTCAAACCATACCACACAAATATCCTTGGCATCGGCCCTGAATTCCTTCTGCCGCTCTGCTACCCGGTGCTTCTGTCCGAAGCAGGGGTTGCTAGGGATCGCTCCGAGTCCGCAAACCCTTGTCGCGTGGCGGGGCTTGTTCGAGCTT gaagagc pSZ4198 (BnLPAT2)SEQ ID NO: 88 gctcttccgctAACGGAGGTCTGTCACCAAATGGACCCCGTCTATTGCGGGAAACCACGGCGATGGCACGTTTCAAAACTTGATGAAATACAATATTCAGTATGTCGCGGGCGGCGACGGCGGGGAGCTGATGTCGCGCTGGGTATTGCTTAATCGCCAGCTTCGCCCCCGTCTTGGCGCGAGGCGTGAACAAGCCGACCGATGTGCACGAGCAAATCCTGACACTAGAAGGGCTGACTCGCCCGGCACGGCTGAATTACACAGGCTTGCAAAAATACCAGAATTTGCACGCACCGTATTCGCGGTATTTTGTTGGACAGTGAATAGCGATGCGGCAATGGCTTGTGGCGTTAGAAGGTGCGACGAAGGTGGTGCCACCACTGTGCCAGCCAGTCCTGGCGGCTCCCAGGGCCCCGATCAAGAGCCAGGACATCCAAACTACCCACAGCATCAACGCCCCGGCCTATACTCGAACCCCACTTGCACTCTGCAATGGTATGGGAACCACGGGGCAGTCTTGTGTGGGTCGCGCCTATCGCGGTCGGCGAA GACCGGGAAggtaccgcggtgagaatcgaaaatgcatcgtttctaggttcggagacggtcaattccctgctccggcgaatctgtcggtcaagctggccagtggacaatgttgctatggcagcccgcgcacatgggcctcccgacgcggccatcaggagcccaaacagcgtgtcagggtatgtgaaactcaagaggtccctgctgggcactccggccccactccgggggcgggacgccaggcattcgcggtcggtcccgcgcgacgagcgaaatgatgattcggttacgagaccaggacgtcgtcgaggtcgagaggcagcctcggacacgtctcgctagggcaacgccccgagtccccgcgagggccgtaaacattgtttctgggtgtcggagtgggcattttgggcccgatccaatcgcctcatgccgctctcgtctggtcctcacgttcgcgtacggcctggatcccggaaagggcggatgcacgtggtgttgccccgccattggcgcccacgtttcaaagtccccggccagaaatgcacaggaccggcccggctcgcacaggccatgctgaacgcccagatttcgacagcaacaccatctagaataatcgcaaccatccgcgttttgaacgaaacgaaacggcgctgtttagcatgtttccgacatcgtgggggccgaagcatgctccggggggaggaaagcgtggcacagcggtagcccattctgtgccacacgccgacgaggaccaatccccggcatcagccttcatcgacggctgcgccgcacatataaagccggacgcctaaccggtttcgtggttatgactagtATGttcgcgttctacttcctgacggcctgcatctccctgaagggcgtgacggcgtctccccctcctacaacggcctgggcctgacgccccagatgggctgggacaactggaacacgacgcctgcgacgtctccgagcagctgctgctggacacggccgaccgcatctccgacctgggcctgaaggacatgggctacaagtacatcatcctggacgactgctggtcctccggccgcgactccgacggcacctggtcgccgacgagcagaagaccccaacggcatgggccacgtcgccgaccacctgcacaacaactccacctgacggcatgtactcctccgcgggcgagtacacgtgcgccggctaccccggctccctgggccgcgaggaggaggacgcccagacttcgcgaacaaccgcgtggactacctgaagtacgacaactgctacaacaagggccagacggcacgcccgagatctcctaccaccgctacaaggccatgtccgacgccctgaacaagacgggccgccccatcactactccctgtgcaactggggccaggacctgaccactactggggctccggcatcgcgaactcctggcgcatgtccggcgacgtcacggcggagacacgcgccccgactcccgctgcccctgcgacggcgacgagtacgactgcaagtacgccggcaccactgctccatcatgaacatcctgaacaaggccgcccccatgggccagaacgcgggcgtcggcggctggaacgacctggacaacctggaggtcggcgtcggcaacctgacggacgacgaggagaaggcgcacactccatgtgggccatggtgaagtcccccctgatcatcggcgcgaacgtgaacaacctgaaggcctcctcctactccatctactcccaggcgtccgtcatcgccatcaaccaggactccaacggcatccccgccacgcgcgtctggcgctactacgtgtccgacacggacgagtacggccagggcgagatccagatgtggtccggccccctggacaacggcgaccaggtcgtggcgctgctgaacggcggctccgtgtcccgccccatgaacacgaccctggaggagatcacttcgactccaacctgggctccaagaagctgacctccacctgggacatctacgacctgtgggcgaaccgcgtcgacaactccacggcgtccgccatcctgggccgcaacaagaccgccaccggcatcctgtacaacgccaccgagcagtcctacaaggacggcctgtccaagaacgacacccgcctgacggccagaagatcggctccctgtcccccaacgcgatcctgaacacgaccgtccccgcccacggcatcgcgactaccgcctgcgcccctcctccTGAtacgtactcgagGCAGCAGCAGCTCGGATAGTATCGACACACTCTGGACGCTGGTCGTGTGATGGACTGTTGCCGCCACACTTGCTGCCTTGACCTGTGAATATCCCTGCCGCTTTTATCAAACAGCCTCAGTGTGTTTGATCTTGTGTGTACGCGCTTTTGCGAGTTGCTAGCTGCTTGTGCTATTTGCGAATACCACCCCCAGCATCCCCTTCCCTCGTTTCATATCGCTTGCATCCCAACCGCAACTTATCTACGCTGTCCTGCTATCCCTCAGCGCTGCTCCTGCTCCTGCTCACTGCCCCTCGCACAGCCTTGGTTTGGGCTCCGCCTGTATTCTCCTGGTACTGCAACCTGTAAACCAGCACTGCAATGCTGATGCACGGGAAGTAGTGGGATGGGAACACAAATGGAAagctgtagaattcctggctcgggcctcgtgctggcactccctcccatgccgacaacctttctgctgtcaccacgacccacgatgcaacgcgacacgacccggtgggactgatcggttcactgcacctgcatgcaattgtcacaagcgcatactccaatcgtatccgtttgatttctgtgaaaactcgctcgaccgcccgcgtcccgcaggcagcgatgacgtgtgcgtgacctgggtgtttcgtcgaaaggccagcaaccccaaatcgcaggcgatccggagattgggatctgatccgagcttggaccagatcccccacgatgcggcacgggaactgcatcgactcggcgcggaacccagctttcgtaaatgccagattggtgtccgataccttgatttgccatcagcgaaacaagacttcagcagcgagcgtatttggcgggcgtgctaccagggttgcatacattgcccatttctgtctggaccgctttaccggcgcagagggtgagttgatggggttggcaggcatcgaaacgcgcgtgcatggtgtgtgtgtctgttttcggctgcacaatttcaatagtcggatgggcgacggtagaattgggtgttgcgctcgcgtgcatgcctcgccccgtcgggtgtcatgaccgggactggaatcccccctcgcgaccctcctgctaacgctcccgactctcccgcccgcgcgcaggatagactctagttcaaccaatcgacaactagtATGgccatggccgccgccgtgatcgtgcccctgggcatcctgttcttcatctccggcctggtggtgaacctgctgcaggccatctgctacgtgctgatccgccccctgtccaagaacacctaccgcaagatcaaccgcgtggtggccgagaccctgtggctggagctggtgtggatcgtggactggtgggccggcgtgaagatccaggtgacgccgacaacgagaccttcaaccgcatgggcaaggagcacgccctggtggtgtgcaaccaccgctccgacatcgactggctggtgggctggatcctggcccagcgctccggctgcctgggctccgccctggccgtgatgaagaagtcctccaagacctgcccgtgatcggctggtccatgtggactccgagtacctgacctggagcgcaactgggccaaggacgagtccaccctgaagtccggcctgcagcgcctgaacgacttcccccgccccactggctggccctgacgtggagggcacccgatcaccgaggccaagctgaaggccgcccaggagtacgccgcctcctccgagctgcccgtgccccgcaacgtgctgatcccccgcaccaagggatcgtgtccgccgtgtccaacatgcgctccacgtgcccgccatctacgacatgaccgtggccatccccaagacctcccccccccccaccatgctgcgcctgacaagggccagccctccgtggtgcacgtgcacatcaagtgccactccatgaaggacctgcccgagtccgacgacgccatcgcccagtggtgccgcgaccagacgtggccaaggacgccctgctggacaagcacatcgccgccgacaccaccccggccagcaggagcagaacatcggccgccccatcaagtccctggccgtggtgctgtcctggtcctgcctgctgatcctgggcgccatgaagacctgcactggtccaacctgactcctcctggaagggcatcgccactccgccctgggcctgggcatcatcaccctgtgcatgcagatcctgatccgctcctcccagtccgagcgctccacccccgccaaggtggtgcccgccaagcccaaggacaaccacaacgactccggctcctcctcccagaccgaggtggagaagcagaagTGA atcgatagatctcttaagGCAGCAGCAGCTCGGATAGTATCGACACACTCTGGACGCTGGTCGTGTGATGGACTGTTGCCGCCACACTTGCTGCCTTGACCTGTGAATATCCCTGCCGCTTTTATCAAACAGCCTCAGTGTGTTTGATCTTGTGTGTACGCGCTTTTGCGAGTTGCTAGCTGCTTGTGCTATTTGCGAATACCACCCCCAGCATCCCCTTCCCTCGTTTCATATCGCTTGCATCCCAACCGCAACTTATCTACGCTGTCCTGCTATCCCTCAGCGCTGCTCCTGCTCCTGCTCACTGCCCCTCGCACAGCCTTGGTTTGGGCTCCGCCTGTATTCTCCTGGTACTGCAACCTGTAAACCAGCACTGCAATGCTGATGCACGGGAAGTAGTGGGATGGGAACACAAATGGAaagettaattaagagatc AGCGGCGACGGTCCTGCTACCGTACGACGTTGGGCACGCCCATGAAAGTTTGTATACCGAGCTTGTTGAGCGAACTGCAAGCGCGGCTCAAGGATACTTGAACTCCTGGATTGATATCGGTCCAATAATGGATGGAAAATCCGAACCTCGTGCAAGAACTGAGCAAACCTCGTTACATGGATGCACAGTCGCCAGTCCAATGAACATTGAAGTGAGCGAACTGTTCGCTTCGGTGGCAGTACTACTCAAAGAATGAGCTGCTGTTAAAAATGCACTCTCGTTCTCTCAAGTGAGTGGCAGATGAGTGCTCACGCCTTGCACTTCGCTGCCCGTGTCATGCCCTGCGCCCCAAAATTTGAAAAAAGGGATGAGATTATTGGGCAATGGACGACGTCGTCGCTCCGGGAGTCAGGACCGGCGGAAAATAAGAGGCAACACACTC CGCTTCTTA gctcttcpSZ4198 BnLPAT2(1.5) SEQ ID NO: 89ATGgccatggccgccgccgccgtgatcgtgcccctgggcatcctgacttcatctccggcctggtggtgaacctgctgcaggccgtgtgctacgtgctgatccgccccctgtccaagaacacctaccgcaagatcaaccgcgtggtggccgagaccctgtggctggagctggtgtggatcgtggactggtgggccggcgtgaagatccaggtgacgccgacgacgagaccacaaccgcatgggcaaggagcacgccctggtggtgtgcaaccaccgctccgacatcgactggctggtgggctggatcctggcccagcgctccggctgcctgggctccgccctggccgtgatgaagaagtcctccaagacctgcccgtgatcggctggtccatgtggactccgagtacctgacctggagcgcaactgggccaaggacgagtccaccctgaagtccggcctgcagcgcctgaacgacacccccgccccactggctggccctgacgtggagggcacccgatcaccgaggccaagctgaaggccgcccaggagtacgccgcctcctcccagctgcccgtgccccgcaacgtgctgatcccccgcaccaagggcttcgtgtccgccgtgtccaacatgcgctccttcgtgcccgccatctacgacatgaccgtggccatccccaagacctcccccccccccaccatgctgcgcctgacaagggccagccctccgtggtgcacgtgcacatcaagtgccactccatgaaggacctgcccgagtccgacgacgccatcgcccagtggtgccgcgaccagacgtggccaaggacgccctgctggacaagcacatcgccgccgacaccaccccggccagaaggagcacaacatcggccgccccatcaagtccctggccgtggtggtgtcctgggcctgcctgctgaccctgggcgccatgaagacctgcactggtccaacctgactcctccctgaagggcatcgccctgtccgccctgggcctgggcatcatcaccctgtgcatgcagatcctgatccgctcctcccagtccgagcgctccacccccgccaaggtggcccccgccaagcccaaggacaagcaccagtccggctcctcctcccagaccgaggtggaggagaagcagaagTGApSZ4206 TcLPAT2 GhomLPAT2A SEQ ID NO: 90ATGgccatcgccgccgccgccgtgatcgtgcccctgggcctgctgttcttcatctccggcctggtggtgaacctgatccaggccctgtgcttcgtgctgatccgccccctgtccaagaacacctaccgcaagatcaaccgcgtggtggccgagctgctgtggctggagctgatctggctggtggactggtgggccggcgtgaagatcaaggtgttcatggaccccgagtccttcaacctgatgggcaaggagcacgccctggtggtggccaaccaccgctccgacatcgactggctggtgggctggctgctggcccagcgctccggctgcctgggctccgccctggccgtgatgaagaagtcctccaagttcctgcccgtgatcggctggtccatgtggttctccgagtacctgttcctggagcgctcctgggccaaggacgagaacaccctgaaggccggcctgcagcgcctgaaggacttcccccgccccttctggctggccttcttcgtggagggcacccgcttcacccaggccaagttcctggccgcccaggagtacgccgcctcccagggcctgcccatcccccgcaacgtgctgatcccccgcaccaagggcttcgtgtccgccgtgtcccacatgcgctccttcgtgcccgccatctacgacatgaccgtggccatccccaagtcctccccctcccccaccatgctgcgcctgttcaagggccagccctccgtggtgcacgtgcacatcaagcgctgcctgatgaaggagctgcccgagaccgacgaggccgtggcccagtggtgcaaggacatgttcgtggagaaggacaagctgctggacaagcacatcgccgaggacaccttctccgaccagcccatgcaggacctgggccgccccatcaagtccctgctggtggtggcctcctgggcctgcctgatggcctacggcgccctgaagttcctgcagtgctcctccctgctgtcctcctggaagggcatcgccttcttcctggtgggcctggccatcgtgaccatcctgatgcacatcctgatcctgttctcccagtccgagcgctccacccccgccaaggtggcccccggcaagcccaagaacgacggcgagacctccgaggcccgccgcgacaagcagcagTGANucleotide sequence of the GhomLPAT2A coding sequence, used in thetransforming DNA from pSZ4412. SEQ ID NO: 91ATGgccatccccgccgccatcgtgatcgtgcccgtgggcctgctgttcttcatctccggcctgatcgtgaacctgctgcaggccctgtgcttcgtgctgatccgccccctgtccaagtccgcctaccgcaccatcaaccgccagctggtggagctgctgtggctggagctggtgtgcatcgtggactggtgggcccgcgtgaagatccagctgttcaccgacaaggagaccctgaactccatgggcaaggagcacgccctggtgatgtgcaaccaccgctccgacatcgactggctggtgggctggatcctggcccagcgctccggctgcctgggctccaccgtggccgtgatgaagaagtcctccaaggtgctgcccgtgatcggctggtccatgtggttctccgagtacctgttcctggagcgcaactgggccaaggacgagtccaccctgaagtccggcctgcagcgcctgcgcgacttcccccgccccttctggctggccctgttcgtggagggcacccgcttcacccagcccaagctgctggccgcccaggagtacgccgcctccaccggcctgcccatcccccgcaacgtgctgatcccccgcaccaagggcttcgtgtccgccgtgtccatcacccgctccttcgtgcccgtgatctacgacatcaccgtggccatccccaagtcctccccccagcccaccatgctgcgcctgttcaagggccagtcctccgtggtgcacgtgcacctgaagcgccacctgatgaaggacctgcccgagtccgacgacgacgtggcccagtggtgccgcgaccagttcgtggtgaaggactccctgctggacaagcacatcgccgaggacaccttctccgaccaggagctgcaggacatcggccgccccatcaagtccctggtggtgttcacctcctgggtgtgcatcatcaccttcggcgccctgaagttcctgcagtggtcctccctgctgcactcctggaagggcatcgccatctccgcctccggcctggccatcgtgaccgtgctgatgcacatcctgatccgcttctcccagtccgagcactccacctccgccaagatcgccgccgagaagcacaagaacggcggcgtgtcccaggagatgggccgcgagaagcagcacTGANucleotide sequence of the GhomLPAT2B coding sequence, used in thetransforming DNA from pSZ4413. SEQ ID NO: 92ATGgagatccccgccgtggccgtgatcgtgcccatcggcatcctgttcttcatctccggcctgatcgtgaacctgatgcaggccatctgcttcttcctgatccgccccctgtccaagaacacccaccgcatcgtgaaccgccagctggccgagctgctgtggctggagctgatctggatcgtggactggtgggccggcgtgaagatccagctgttcaccgacaaggagaccctgcacctgatgggcaaggagcacgccctggtgatctgcaaccactcctccgacatcgactggctggtgggctggctgctgtgccagcgctccggctgcctgggctccgccctggccgtgatgaagtcctcctccaaggtgctgcccgtgatcggctggtccatgtggttctccgagtacctgttcctggagcgctcctgggccaaggacgagtccaccctgaagtccggcctgcagcgcctgaaggacttcccccgccccttctggctggccctgttcgtggagggcacccgcttcacccaggccaagctgctggccgcccaggagtacgccatgtccgccggcctgcccgtgccccgcaacgtgctgatcccccgcaccaagggcttcgtgtccgccgtgtccaacatgcgctccttcgtgcccgccatctacgacgtgaccgtggccatccccaagtcctccgtgcagcccaccatgctgcgcctgttcaagggccagtcctccgtggtgcaggtgcacctgaagcgccactccatgaaggacctgcccgagtccgaggacgacgtggcccagtggtgccgcgaccgcttcgtggtgaaggactccctgctggacaagcacaaggtggaggacaccttcaccgaccaggagctgcaggacctgggccgccccatcaagtccctggtggtggtgacctgctgggcctgcatcatcatcttcggcatcctgaagttcctgcagtggtcctccctgctgtactcctggaagggcatggccatctccgcctccggcctggccgtggtgaccttcctgatgcagatcctgatccgcttctcccagtccgagcgctccacccccgccaagatcgcccccgccaagcccaacaaggccggcaactcctccgagaccgtgcgcgacaagcaccagTGANucleotide sequence of the GhomLPAt2C coding sequence, used in thetransforming DNA from pSZ4414. SEQ ID NO: 93ATGgccatccccgccgccatcatcatcgtgcccctgggcctgatcttcttcacctccggcctgatcatcaacctgatccaggccgtgtgctacgtgctgatccgccccctgtccaagtccaccttccgccgcatcaaccgcgagctggccgagctgctgtggctggagctggtgtgggtggtggactggtgggccggcgtgaagatccagctgttcaccgacaaggagaccctgcactccatgggcaaggagcacgccctggtgatctgcaaccaccgctccgacatcgactggctggtgggctggatcctggcccagcgctccggctgcctgggctccgccctggccgtgatgaagaagtcctccaaggtgctgcccgtgatcggctggtccatgtggttctccgagtacttcttcctggagcgcaactgggccatggacgagtccaccctgaagtccggcctgcagcgcctgaaggacttcccccagcccttctggctggccctgttcgtggagggcacccgcttcacccagcccaagctgctggccgcccaggagtacgccgcctccgccggcctgcccatcccccgcaacgtgctgatcccccgcaccaagggcttcgtgtccgccgtgaacatcatgcgctccttcgtgcccgccatctacgacgtgaccgtggccatccccaagtcctccccccagcccaccatgctgcgcctgttcaagggccagtcctccgtggtgcacgtgcacctgaagcgccacctgatggaggacctgcccgagaccgacgacgacgtggcccagtggtgccgcgaccgcttcgtggtgaaggactccctgctggacaagtacgtggccgaggacaccttctccgaccaggagctgcaggacctgggccgccccatcaagtccctggtggtggtgacctcctgggtgtgcatcatcgccttcggctccctgaagttcctgcagtggtcctccctgctgtactcctggaagggcatcgtgatctccgccgcctccctggccgtggtgaccgtgctgatgcagatcctgatccgcttctcccagtccgagcgctccacctccgccaagatcgccgccgccaagcgcaagaacgtgggcgagcacTGANucleotide sequence of the GindPAT2A coding sequence, used in the transformingDNA from pSZ4415. SEQ ID NO: 94ATGgccatccccgtggtggtggtgatcgtgcccgtgggcctgctgttcttcatctccggcctgatcgtgaacctgctgcaggccctgtgcttcgtgctgatccgccccctgtccaagtccgcctaccgcaccatcaaccgccagctggtggagctgctgtggctggagctggtgtgcatcgtggactggtgggcccgcgtgaagatccagctgttcatcgacaaggagaccctgaactccatgggcaaggagcacgccctggtgatgtgcaaccaccgctcctacatcgactggctggtgggctggatcctggcccagcgctccggctgcctgggctccaccgtggccgtgatgaagaagtcctccaaggtgctgcccgtgatcggctggtccatgtggttctccgagtacctgttcctggagcgcaactgggccaaggacgagtccaccctgaagtccggcctgcagcgcctgcgcgacttcccccgccccttctggctggccctgttcgtggagggcacccgcttcacccagcccaagctgctggccgcccaggagtacgccgcctccaccggcctgcccatcccccgcaacgtgctgatcccccgcaccaagggcttcgtgtccgccgtgtccatcacccgctccttcgtgcccgtgatctacgacatcaccgtggccatccccaagtcctcctcccagcccaccatgctgaagctgttcaagggccagtcctccgtggtgcacgtgcacctgaagcgccacctgatgaaggacctgcccgagtccgacgacgacgtggcccagtggtgccgcgcccagttcgtggtgaaggactccctgctggacaagcacatcgccgaggacaccttctccgaccaggagctgcaggacatcggccgccccatcaagtccctggtggtgttcacctcctgggtgtgcatcatcaccttcggcgccctgaagttcctgcagtggtcctccctgctgcactcctggaagggcatcgccatctccgcctccggcctggccatcgtgaccgtgctgatgcacatcctgatccgcttctcccagtccgagcactccacctccgccaagatcgccgccgagaagcacaagaacggcggcgtgtcccaggagatgggccgcgagaagcagcacTGANucleotide sequence of the GindPAT2B coding sequence, used in the transofrmingDNA from pSZ4416. SEQ ID NO: 95ATGggcatccccgccgtggccgtgatcgtgcccatcggcatcctgttcttcatctccggcttcatcgtgaacctgatgcaggccatctgcttcgtgctgatccgccccctgtccaagaacacctaccgcatcgtgaaccgccagctggccgagttcctgtggctggagctgatctgggtggtggactggtgggccggcgtgaagatccagctgttcaccgacaaggagaccctgcacctgatgggcaaggagcacgccctggtgatctgcaaccaccgctccgacatcgactggctggtgggctggctgctgtgccagcgctccggctgcctgggctccgccctggccgtgatgaagtcctcctccaaggtgctgcccgtgatcggctggtccatgtggttctccgagtacctgttcctggagcgctcctgggccaaggacgagtccaccctgaagctgggcctgcagcgcctgaaggacttcccccgccccttctggctggccctgttcgtggagggcacccgcttcacccaggccaagctgctggccgcccaggagtacgccatgtccgccggcctgcccgtgccccgcaacgtgctgatcccccgcaccaagggcttcgtgtccgccgtgtccaacatgcgctccttcgtgcccgccatctacgacgtgaccgtggccatccccaagtcctccgtgcagcccaccatgctgggcctgttcaagggccagtcctgcgtggtgcaggtgcacctgaagcgccacctgatgaaggacctgcccgagtccgaggacgacgtggcccagtggtgccgcgagcgcttcgtggtgaaggactccctgctggacaagcacaaggtggaggacaccttctccgaccaggagctgcaggacctgggccgccccatcaagtccctggtggtggtgatctcctgggcctgcatcctgatcttctggatcctgaagttcctgcagtggtcctccctgctgtactcctggaagggcatcgccatctccgcctgcgccatggccgtgatcgccttcctgatgcagatcctgctgcgcttctcccagtccgagcgctccacccccgccaagatcgcccccgccaagcccaacaacgcccgcaactcctccgagaccgtgcgcgacaagcaccagTGANucleotide sequence of the GindPAT2C coding sequence, used in the transformingDNA from pSZ4417. SEQ ID NO: 96ATGgccatccccgccgccatcatcatcgtgcccctgggcctgatcttcttcacctccggcttcatcatcaacctgatccaggccgtgtgctacgtgctgatccgccccctgtccaagtccaccttccgccgcatcaaccgccagctggccgagctgctgtggctggagctggtgtgggtggtggactggtgggccggcgtgaagatccagctgttcaccaacaaggagaccctgcactccatcggcaaggagcacgccctggtgatctgcaaccagcgctccgacatcgactggctggtgggctggatcctggcccagcgctccggctgcctgggctccgccctggccgtgatgaagaagtcctccaaggtgctgcccgtgatcggctggtccatgtggttctccgagtacctgttcctggagcgcaactgggccatggacgagtccaccctgaagtccggcctgcagtggctgaaggacttcccccagcccttctggctggccctgttcgtggagggcacccgcttcacccagcccaagctgctggccgcccaggagtacgccgcctccgccggcctgcccatcccccgcaacgtgctgatcccccgcaccaagggcttcgtgtccgccgtgaacatcatgcgctccttcgtgcccgccgtgtacgacgtgaccgtggccatccccaagtcctccccccagcccaccatgctgcgcctgttcaagggccagtcctccgtggtgcacgtgcacctgaagcgccacctgatggaggacctgcccgagaccgacgacgacgtggcccagtggtgccgcgaccgcttcgtggtgaaggactccctgctggacaagcacctggccgaggacaccttctccgaccaggagctgcaggacctgggccgccccatcaagtccctggtggtggtgacctcctgggtgtgcatcatcgccttcggcgccctgaagttcctgcagtggtcctccctgctgtactcctggaagggcatcgtgatctccgccgcctccctggccgtggtgaccgtgctgatgcagatcctgatccgcttctcccagtccgagcgctccacctccgccaaggtggtggccgagaagcgcaagaacgtgggcgagcacTGApSZEX61 Transorming DNA expressing CnLPAAT. SEQ ID NO: 97gtttaaacgccggtcaccacccgcatgctcgtactacagcgcacgcaccgcttcgtgatccaccgggtgaacgtagtcctcgacggaaacatctggttcgggcctcctgcttgcactcccgcccatgccgacaacctttctgctgttaccacgacccacaatgcaacgcgacacgaccgtgtgggactgatcggttcactgcacctgcatgcaattgtcacaagcgcttactccaattgtattcgtttgttttctgggagcagttgctcgaccgcccgcgtcccgcaggcagcgatgacgtgtgcgtggcctgggtgtttcgtcgaaaggccagcaaccctaaatcgcaggcgatccggagattgggatctgatccgagtttggaccagatccgccccgatgcggcacgggaactgcatcgactcggcgcggaacccagctttcgtaaatgccagattggtgtccgatacctggatttgccatcagcgaaacaagacttcagcagcgagcgtatttggcgggcgtgctaccagggttgcatacattgcccatttctgtctggaccgctttactggcgcagagggtgagttgatggggttggcaggcatcgaaacgcgcgtgcatggtgtgcgtgtctgttttcggctgcacgaattcaatagtcggatgggcgacggtagaattgggtgtggcgctcgcgtgcatgcctcgccccgtcgggtgtcatgaccgggactggaatcccccctcgcgaccatcttgctaacgctcccgactctcccgaccgcgcgcaggatagactcttgttcaaccaatcgaca ggtaccATGgacgcctccggcgcctcctccttcctgcgcggccgctgcctggagtcctgcttcaaggcctccttcggctacgtaatgtcccagcccaaggacgccgccggccagccctcccgccgccccgccgacgccgacgacttcgtggacgacgaccgctggatcaccgtgatcctgtccgtggtgcgcatcgccgcctgcttcctgtccatgatggtgaccaccatcgtgtggaacatgatcatgctgatcctgctgccctggccctacgcccgcatccgccagggcaacctgtacggccacgtgaccggccgcatgctgatgtggattctgggcaaccccatcaccatcgagggctccgagttctccaacacccgcgccatctacatctgcaaccacgcctccctggtggacatcttcctgatcatgtggctgatccccaagggcaccgtgaccatcgccaagaaggagatcatctggtatcccctgttcggccagctgtacgtgctggccaaccaccagcgcatcgaccgctccaacccctccgccgccatcgagtccatcaaggaggtggcccgcgccgtggtgaagaagaacctgtccctgatcatcttccccgagggcacccgctccaagaccggccgcctgctgcccttcaagaagggcttcatccacatcgccctccagacccgcctgcccatcgtgccgatggtgctgaccggcacccacctggcctggcgcaagaactccctgcgcgtgcgccccgcccccatcaccgtgaagtacttctcccccatcaagaccgacgactgggaggaggagaagatcaaccactacgtggagatgatccacgccctgtacgtggaccacctgcccgagtcccagaagcccctggtgtccaagggccgcgacgcctccggccgctccaactccTGAttaattaactcgagatgtggagatgtagggtggtcgactcgttggaggtgggtgtttttttttatcgagtgcgcggcgcggcaaacgggtccctttttatcgaggtgttcccaacgccgcaccgccctcttaaaacaacccccaccaccacttgtcgaccttctcgtttgttatccgccacggcgccccggaggggcgtcgtctggccgcgcgggcagctgtatcgccgcgctcgctccaatggtgtgtaatcttggaaagataataatcgatggatgaggaggagagcgtgggagatcagagcaaggaatatacagttggcacgaagcagcagcgtactaagctgtagcgtgttaagaaagaaaaactcg

cgtctccccctcctacaacggcctgggcctgacgccccagatgggctgggacaactggaacacgttcgcctgcgacgtctccgagcagctgctgctggacacggccgaccgcatctccgacctgggcctgaaggacatgggctacaagtacatcatcctggacgactgctggtcctccggccgcgactccgacggcttcctggtcgccgacgagcagaagaccccaacggcatgggccacgtcgccgaccacctgcacaacaactccttcctgacggcatgtactcctccgcgggcgagtacacgtgcgccggctaccccggctccctgggccgcgaggaggaggacgcccagttcttcgcgaacaaccgcgtggactacctgaagtacgacaactgctacaacaagggccagttcggcacgcccgagatctcctaccaccgctacaaggccatgtccgacgccctgaacaagacgggccgccccatcttctactccctgtgcaactggggccaggacctgaccttctactggggctccggcatcgcgaactcctggcgcatgtccggcgacgtcacggcggagttcacgcgccccgactcccgctgcccctgcgacggcgacgagtacgactgcaagtacgccggcttccactgctccatcatgaacatcctgaacaaggccgcccccatgggccagaacgcgggcgtcggcggctggaacgacctggacaacctggaggtcggcgtcggcaacctgacggacgacgaggagaaggcgcacttctccatgtgggccatggtgaagtcccccctgatcatcggcgcgaacgtgaacaacctgaaggcctcctcctactccatctactcccaggcgtccgtcatcgccatcaaccaggactccaacggcatccccgccacgcgcgtctggcgctactacgtgtccgacacggacgagtacggccagggcgagatccagatgtggtccggccccctggacaacggcgaccaggtcgtggcgctgctgaacggcggctccgtgtcccgccccatgaacacgaccctggaggagatcttcttcgactccaacctgggctccaagaagctgacctccacctgggacatctacgacctgtgggcgaaccgcgtcgacaactccacggcgtccgccatcctgggccgcaacaagaccgccaccggcatcctgtacaacgccaccgagcagtcctacaaggacggcctgtccaagaacgacacccgcctgttcggccagaagatcggctccctgtcccccaacgcgatcctgaacacgaccgtccccgcccacggcatcgcgactaccgcctgcgcccctcctccTGA tacaacttattacgtattctgaccggcgctgatgtggcgcggacgccgtcgtactctttcagactttactcttgaggaattgaacctttctcgcttgctggcatgtaaacattggcgcaattaattgtgtgatgaagaaagggtggcacaagatggatcgcgaatgtacgagatcgacaacgatggtgattgttatgaggggccaaacctggctcaatcttgtcgcatgtccggcgcaatgtgatccagcggcgtgactctcgcaacctggtagtgtgtgcgcaccgggtcgctttgattaaaactgatcgcattgccatcccgtcaactcacaagcctactctagctcccattgcgcactcgggcgcccggctcgatcaatgttctgagcggagggcgaagcgtcaggaaatcgtctcggcagctggaagcgcatggaatgcggagcggagatcgaatcagatatcAAGCTCCATC gagctc cagccacggcaacaccgcgcgccttgcggccgagcacggcgacaagaacctgagcaagatctgcgggctgatcgccagcgacgagggccggcacgagatcgcctacacgcgcatcgtggacgagttcttccgcctcgaccccgagggcgccgtcgccgcctacgccaacatgatgcgcaagcagatcaccatgcccgcgcacctcatggacgacatgggccacggcgaggccaacccgggccgcaacctcttcgccgacttctccgcggtcgccgagaagatcgacgtctacgacgccgaggactactgccgcatcctggagcacctcaacgcgcgctggaaggtggacgagcgccaggtcagcggccaggccgccgcggaccaggagtacgtcctgggcctgccccagcgcttccggaaactcgccgagaagaccgccgccaagcgcaagcgcgtcgcgcgcaggcccgtcgccttctcctggatctccgggcgcgagatcatggtctagggagcgacgagtgtgcgtgcggggctggcgggagtgggacgccctcctcgctcctctctgttctgaacggaacaatcggccaccccgcgctacgcgccacgcatcgagcaacgaagaaaaccccccgatgataggttgcggtggctgccgggatatagatccggccgcacatcaaagggcccctccgccagagaagaagctcctttcccagcagactcct gaagagcgtttaaacCpauLPAAT1 SEQ ID NO: 98 ggtaccATGgccatccccgccgccgccgtgatcttcctgttcggcctgctgttcttcacctccggacctgatcatcaacctgttccaggccctgtgcttcgtgctggtgtggcccctgtccaagaacgcctaccgccgcatcaaccgcgtgttcgccgagctgctgctgtccgagctgctgtgcctgttcgactggtgggccggcgccaagctgaagctgttcaccgaccccgagaccttccgcctgatgggcaaggagcacgccctggtgatcatcaaccacatgaccgagctggactggatgctgggctgggtgatgggccagcacctgggctgcctgggctccatcctgtccgtggccaagaagtccaccaagttcctgcccgtgctgggctggtccatgtggttctccgagtacctgtacatcgagcgctcctgggccaaggaccgcaccaccctgaagtcccacatcgagcgcctgaccgactaccccctgcccttctggatggtgatcttcgtggagggcacccgcttcacccgcaccaagctgctggccgcccagcagtacgccgcctcctccggcctgcccgtgccccgcaacgtgctgatcccccgcaccaagggcttcgtgtcctgcgtgtcccacatgcgctccttcgtgcccgccgtgtacgacgtgaccgtggccttccccaagacctcccccccccccaccctgctgaacctgttcgagggccagtccatcgtgctgcacgtgcacatcaagcgccacgccatgaaggacctgcccgagtccgacgacgccgtggcccagtggtgccgcgacaagttcgtggagaaggacgccctgctggacaagcacaacgccgaggacaccttctccggccaggaggtgcaccgcaccggctcccgccccatcaagtccctgctggtggtgatctcctgggtggtggtgatcaccttcggcgccctgaagttcctgcagtggtcctcctggaagggcaaggccttctccgtgatcggcctgggcatcgtgaccctgctgatgcacatgctgatcctgtcctcccaggccgagcgctcctccaaccccgccaaggtggcccaggccaagctgaagaccgagctgtccatctccaagaaggccaccgacaaggagaacTGA ctcgagCprocLPAAT1 SEQ ID NO: 99ggtaccATGgccatccccgccgccgccgtgatcttcctgttcggcctgatcttcttcgcctccggcctgatcatcaacctgttccaggccctgtgcttcgtgctgatctggcccatctccaagaacgcctaccgccgcatcaaccgcgtgttcgccgagctgctgctgtccgagctgctgtgcctgttcgactggtgggccggcgccaagctgaagctgttcaccgaccccgagaccttccgcctgatgggcaaggagcacgccaggtgatcatcaaccacatgaccgagaggactggatggtgggagggtgatgggccagcactteggagcctgggctccatcctgtccgtggccaagaagtccaccaagttcctgcccgtgctgggctggtccatgtggttcaccgagtacctgtacatcgagcgctcctggaacaaggacaagtccaccctgaagtcccacatcgagcgcctgaaggactaccccctgcccttctggctggtgatcttcgccgagggcacccgatcacccagaccaagagaggccgcccagcagtacgccgcctcctccggcctgcccgtgccccgcaacgtgctgatcccccgcaccaagggcttcgtgtcctgcgtgtcccacatgcgctccttcgtgcccgccgtgtacgacctgaccgtggccttccccaagacctcccccccccccaccctgctgaacctgttcgagggccagtccgtggtgctgcacgtgcacatcaagcgccacgccatgaaggacctgcccgagtccgacgacgaggtggcccagtggtgccgcgacaagncgtggagaaggacgccagaggacaagcacaacgccgaggacaccttaccggccaggagagcagcacaccggccgccgccccatcaagtccctgaggtggtgatctcctgggtggtggtgatcgccttcggcgccctgaagttcctgcagtggtcctcctggaagggcaaggccttctccgtgatcggcctgggcatcgtgaccagagatgcacatgagatcctgtecteccaggccgagcgaccaagcccgccaaggtggcccaggccaagctgaagaccgagctgtccatctccaagaccgtgaccgacaaggagaacTGActcgag CpaiLPAAT1SEQ ID NO: 100 ggtaccATGgccatcccctccgccgccgtggtgttcctgttcggcctgctgttcttcacctccggcctgatcatcaacctgttccaggccttctgcttcgtgctgatctcccccctgtccaagaacgcctaccgccgatcaaccgcgtgttcgccgagctgctgcccctggagttcctgtggctgaccactggtgcgccggcgccaagctgaagctgacaccgaccccgagaccaccgcctgatgggcaaggagcacgccctggtgatcatcaaccacaagatcgagctggactggatggtgggctgggtgctgggccagcacctgggctgcctgggctccatcctgtccgtggccaagaagtccaccaagacctgcccgtgacggctggtccctgtggactccggctacctgacctggagcgctcctgggccaaggacaagatcaccctgaagtcccacatcgagtccctgaaggactaccccctgccataggctgatcatcacgtggagggcacccgatcacccgcaccaagctgctggccgcccagcagtacgccgcctcctccggcctgcccgtgccccgcaacgtgctgatcccccacaccaagggcttcgtgtcctccgtgtcccacatgcgctccttcgtgcccgccatctacgacgtgaccgtggccttccccaagacctcccccccccccaccatgctgaagctgacgagggccagtccgtggagctgcacgtgcacatcaagcgccacgccatgaaggacctgcccgagtccgacgacgccgtggcccagtggtgccgcgacaagacgtggagaaggacgccctgctggacaagcacaactccgaggacaccactccggccaggaggtgcaccacgtgggccgccccatcaaggccctgctggtggtgatctcctgggtggtggtgatcatcacggcgccctgaagacctgctgtggtcctccctgctgtcctcctggaagggcaaggccactccgtgatcggcctgggcatcgtggccggcatcgtgaccctgctgatgcacatcctgatcctgtcctcccaggccgagggctccaaccccgtgaaggccgcccccgccaagctgaagaccgagctgtcctcctccaagaaggtgaccaacaggagaacTGA ctcgagChookLPAAT1 SEQ ID NO: 101 ggtaccATGgccatcccctccgccgccgtggtgttcctgttcggcctgctgttcttcacctccggcctgatcatcaacctgttccaggccttctgcttcgtgctgatctcccccctgtccaagaacgcctaccgccgcatcaaccgcgtgttcgccgagctgctgcccctggagttcctgtggctgaccactggtgcgccggcgccaagctgaagctgacaccgaccccgagaccaccgcctgatgggcaaggagcacgccctggtgatcatcaaccacaagatcgagctggactggatggtgggctgggtgctgggccagcacctgggctgcctgggctccatcctgtccgtggccaagaagtccaccaagacctgcccgtgacggctggtccctgtggactccgagtacctgacctggagcgctcctgggccaaggacaagatcaccctgaagtcccacatcgagtccctgaaggactaccccctgccataggctgatcatcacgtggagggcacccgatcacccgcaccaagctgctggccgcccagcagtacgccgcctcctccggcctgcccgtgccccgcaacgtgctgatcccccacaccaagggcttcgtgtcctccgtgtcccacatgcgctccttcgtgcccgccatctacgacgtgaccgtggccttccccaagacctcccccccccccaccatgctgaagctgacgagggccagtccgtggagctgcacgtgcacatcaagcgccacgccatgaaggacctgcccgagtccgacgacgccgtggcccagtggtgccgcgacaagacgtggagaaggacgccctgctggacaagcacaactccgaggacaccactccggccaggaggtgcaccacgtgggccgccccatcaaggccctgctggtggtgatctcctgggtggtggtgatcatcacggcgccctgaagacctgctgtggtcctccctgctgtcctcctggaagggcaaggccactccgtgatcggcctgggcatcgtggccggcatcgtgaccctgctgatgcacatcctgatcctgtcctcccaggccgagggctccaaccccgtgaaggccgcccccgccaagctgaagaccgagctgtcctcctccaagaaggtgaccaacaaggagaacTGA ctcgagCignLPAAT1 SEQ ID NO: 102 ggtaccATGgccatcgccgccgccgccgtgatcttcctgttcggcctgctgttcttcgcctccggcatcatcatcaacctgttccaggccctgtgcacgtgctgatctggcccctgtccaagaacgtgtaccgccgcatcaaccgcgtgacgccgagctgctgctgatggacctgctgtgcctgaccactggtgggccggcgccaagatcaagctgacaccgaccccgagaccaccgcctgatgggcatggagcacgccctggtgatcatgaaccacaagaccgacctggactggatggtgggctggatcctgggccagcacctgggctgcctgggctccatcctgtccatcgccaagaagtccaccaagacatccccgtgctgggctggtccgtgtggactccgagtacctgacctggagcgctcctgggccaaggacaagtccaccctgaagtcccacatggagaagctgaaggactaccccctgccataggctggtgatcacgtggagggcacccgatcacccgcaccaagctgctggccgcccagcagtacgccgcctcctccggcctgcccgtgccccgcaacgtgctgatcccccacaccaagggcttcgtgtcctgcgtgtccaacatgcgctccttcgtgcccgccgtgtacgacgtgaccgtggccttccccaagtcctcccccccccccaccatgctgaagctgacgagggccagtccatcgtgctgcacgtgcacatcaagcgccacgccctgaaggacctgcccgagtccgacgacgccgtggcccagtggtgccgcgacaagacgtggagaaggacgccctgctggacaagcacaacgccgaggacaccactccggccaggaggtgcaccacatcggccgccccatcaagtccctgctggtggtgatcgcctgggtggtggtgatcatcacggcgccctgaagacctgcagtggtcctccctgctgtccacctggaagggcaaggccactccgtgatcggcctgggcatcgccaccctgctgatgcacatgctgatcctgtcctcccaggccgagcgctccaaccccgccaaggtggccaagTGA ctcgag CavigLPAAT1 SEQ ID NO: 103 ggtaccATGaccatcgcctccgccgccgtggtgttcctgttcggcatcctgctgttcacctccggcctgatcatcaacctgttccaggccttctgctccgtgctggtgtggcccctgtccaagaacgcctaccgccgcatcaaccgcgtgttcgccgagttcctgcccctggagttcctgtggctgttccactggtgggccggcgccaagctgaagctgttcaccgaccccgagaccttccgcctgatgggcaaggagcacgccctggtgatcatcaaccacaagatcgagctggactggatggtgggctgggtgctgggccagcacctgggctgcctgggctccatcctgtccgtggccaagaagtccaccaagacctgcccgtgacggctggtccctgtggactccgagtacctgacctggagcgcaactgggccaaggacaagaagaccctgaagtcccacatcgagcgcctgaaggactaccccctgccataggctgatcatcttcgtggagggcacccgatcacccgcaccaagctgctggccgcccagcagtacgccgcctccgccggcctgcccgtgccccgcaacgtgctgatcccccacaccaagggcttcgtgtcctccgtgtcccacatgcgctccttcgtgcccgccatctacgacgtgaccgtggccttccccaagacctcccccccccccaccatgctgaagctgacgagggccacttcgtggagctgcacgtgcacatcaagcgccacgccatgaaggacctgcccgagtccgaggacgccgtggcccagtggtgccgcgacaagacgtggagaaggacgccctgctggacaagcacaacgccgaggacaccactccggccaggaggtgcaccacgtgggccgccccatcaagtccctgctggtggtgatctcctgggtggtggtgatcatcacggcgccctgaagacctgcagtggtcctccctgctgtcctcctggaagggcatcgccactccgtgatcggcctgggcaccgtggccctgctgatgcagatcctgatcctgtcctcccaggccgagcgctccatccccgccaaggagacccccgccaacctgaagaccgagctgtcctcctccaagaaggtgaccaacaaggagaacTGA ctcgagCavigLPAAT2 SEQ ID NO: 104 ggtaccATGgccatcgccgccgccgccgtgatcgtgcccgtgtccctgctgttcttcgtgtccggcctgatcgtgaacctggtgcaggccgtgtgatcgtgctgatccgccccctgacaagaacacctaccgccgcatcaaccgcgtggtggccgagctgctgtggctggagctggtgtggctgatcgactggtgggccggcgtgaagatcaaggtgacaccgaccacgagaccaccacctgatgggcaaggagcacgccctggtgatctgcaaccacaagtccgacatcgactggctggtgggctgggtgctggcccagcgctccggctgcctgggctccaccctggccgtgatgaagaagtcctccaagacctgcccgtgatcggctggtccatgtggactccgagtacctgacctggagcgcaactgggccaaggacgagtccaccctgaagtccggcctgaaccgcctgaaggactaccccctgccataggctggccctgttcgtggagggcacccgatcacccgcgccaagctgctggccgcccagcagtacgccgcctcctccggcctgcccgtgccccgcaacgtgctgatcccccgcaccaagggcttcgtgtcctccgtgtcccacatgcgctccttcgtgcccgccatctacgacgtgaccgtggccatccccaagacctcccccccccccaccctgctgcgcatgacaagggccagtcctccgtgctgcacgtgcacctgaagcgccaccagatgaacgacctgcccgagtccgacgacgccgtggcccagtggtgccgcgacatcacgtggagaaggacgccctgctggacaagcacaacgccgaggacaccactccggccaggagctgcaggacaccggccgccccatcaagtccctgctgatcgtgatctcctgggccgtgctggtggtgacggcgccgtgaagacctgcagtggtcctccctgctgtcctcctggaagggcctggccactccggcatcggcctgggcgtgatcaccctgctgatgcacatcctgatcctgactcccagtccgagcgctccacccccgccaaggtggcccccgccaagcccaagatcgagggcgagtcctccaagaccgagatggagaaggagcacTGA ctcgagCpalLPAAT1 SEQ ID NO: 105 ggtaccATGgccatcgccgccgccgccgtgatcgtgcccctgggcctgctgttcttcgtgtccggcctgatcgtgaacctggtgcaggccgtgtgcttcgtgctgatccgccccctgtccaagaacacctaccgccgcatcaaccgcgtggtggccgagctgctgtggctggagctggtgtggctgatcgactggtgggccggcgtgaagatcaaggtgttcaccgaccacgagaccctgtccctgatgggcaaggagcacgccctggtgatctgcaaccacaagtccgacatcgactggctggtgggctgggtgctggcccagcgctccggctgcctgggctccaccctggccgtgatgaagaagtcctccaagttcctgcccgtgatcggctggtccatgtggttctccgagtacctgcccgagtccgacgacgccgtggcccagtggtgccgcgacatcttcgtggagaaggacgccctgctggacaagcacaacgccgaggacaccttctccggccaggagctgcaggacaccggccgccccatcaagtccctgctggtggtgatctcctgggccgtgctggtgatcttcggcgccgtgaagttcctgcagtggtcctccctgctgtcctcctggaagggcctggccttctccggcgtgggcctgggcatcatcaccctgctgatgcacatcctgatcctgttctcccagtccgagcgctccacccccgccaaggtggcccccgccaagccaagaaggacggcgagtcctccaagaccgagatcgagaaggagaacgttcctggagcgctcctgggccaaggacgagaacaccctgaagtccggcctgaaccgcctgaaggactaccccctgcccUctggctggccctgttcgtggagggcacccgcttcacccgcgccaagctgctggccgcccagcagtacgccacctcctccggcctgcccgtgccccgcaacgtgctgatcccccgcaccaagggcttcgtgtcctccgtgtcccacatgcgctcatcgtgcccgccatctacgacgtgaccgtggccatccccaagacctcccccccccccaccatgctgcgcatgttcaagggccagtcctccgtgctgcacgtgcacctgaagcgccacctgatgaaggacctTGA ctcgagCuPSR23 LPAAT2 SEQ ID NO: 106 ggtaccATGgccatcgccgccgccgccgtgatcttcctgttcggcctgatcttcttcgcctccggcctgatcatcaacctgttccaggccctgtgcttcgtgctgatccgccccctgtccaagaacgcctaccgccgcatcaaccgcgtgttcgccgagctgctgctgtccgagctgctgtgcctgttcgactggtgggccggcgccaagctgaagctgttcaccgaccccgagaccttccgcctgatgggcaaggagcacgccctggtgatcatcaaccacatgaccgagctggactggatggtgggctgggtgatgggccagcacttcggctgcctgggctccatcatctccgtggccaagaagtccaccaagttcctgcccgtgctgggctggtccatgtggttctccgagtacctgtacctggagcgctcctgggccaaggacaagtccaccctgaagtcccacatcgagcgcctgatcgactaccccctgcccttctggctggtgatcttcgtggagggcacccgcttcacccgcaccaagctgctggccgcccagcagtacgccgtgtcctccggcctgcccgtgccccgcaacgtgctgatcccccgcaccaagggcttcgtgtcctgcgtgtcccacatgcgctcatcgtgcccgccgtgtacgacgtgaccgtggccttccccaagacctcccccccccccaccctgctgaacctgttcgagggccagtccatcatgctgcacgtgcacatcaagcgccacgccatgaaggacctgcccgagtccgacgacgccgtggccgagtggtgccgcgacaagttcgtggagaaggacgccctgctggacaagcacaacgccgaggacaccttctccggccaggaggtgtgccactccggctcccgccagctgaagtccctgctggtggtgatctcctgggtggtggtgaccaccttcggcgccctgaagttcctgcagtggtcctcctggaagggcaaggccttctccgccatcggcctgggcatcgtgaccctgctgatgcacgtgctgatcctgtcctcccaggccgagcgctccaaccccgccgaggtggcccaggccaagctgaagaccggcctgtccatctccaagaaggtgaccgacaaggagaacTGA ctcgag CkoeLPAAT1SEQ ID NO: 107 ggtaccATGgccatccccgccgccgtggccgtgatccccatcggcctgctgttcatcatctccggcctgatcgtgaacctgatccaggccgtggtgtacgtgctgatccgccccctgtccaagaacctgcaccgcaagatcaacaagcccatcgccgagctgctgtggctggagctgatctggctggtggactggtgggccggcatcaaggtggaggtgtacgccgactcccagaccctggagctgatgggcaaggagcacgccctgctgatctgcaaccaccgctccgacatcgactggctggtgggctgggtgctggcccagcgcgcccgctgcctgggctccgccctggccatcatgaagaagtccgccaagttcctgcccgtgatcggctggtccatgtggttctccgactacatcttcctggaccgcacctgggccaaggacgagaagaccctgaagtccggcttcgagcgcctggccgacttccccatgcccttctggctggccctgttcgtggagggcacccgcttcaccaaggccaagctgctggccgcccaggagtacgccgcctcccgcggcctgcccgtgccccagaacgtgctgatcccccgcaccaagggcttcgtgaccgccgtgacccacatgcgctcctacgtgcccgccatctacgactgcaccgtggacatctccaaggcccaccccgccccctccatcctgcgcctgatccgcggccagtcctccgtggtgaaggtgcagatcacccgccactccatgcaggagctgcccgagaccgccgacggcatctcccagtggtgcatggacctgttcgtgaccaaggacggcttcctggagaagtaccactccaaggacatcttcggctccctgcccgtgcagaacatcggccgccccgtgaagtccctgatcgtggtgctgtgctggtactgcctgatggccttcggcctgttcaagttcttcatgtggtcctccctgctgtcctcctgggagggcatcctgtccctgggcctgatcctgctggccgtggccatcgtgatgcagatcctgatccagtccaccgagtccgagcgctccacccccgtgaagtccatccagaaggacccctccaaggagaccctgctgcagaacTGA ctcgag CkoeLPAAT2 SEQ ID NO: 108ggtaccATGcacgtgctgctggagatggtgaccttccgcttctcctccttcttcgtgttcgacaacgtgcaggccctgtgcttcgtgctgatctggcccctgtccaagtccgcctaccgcaagatcaaccgcgtgttcgccgagctgctgctgtccgagctgctgtgcctgttcgactggtgggccggcgccaagctgaagctgttcaccgaccccgagaccttccgcctgatgggcaaggagcacgccctggtgatcaccaaccacaagatcgacctggactggatgatcggctggatcctgggccagcacttcggctgcctgggctccgtgatctccatcgccaagaagtccaccaagttcctgcccatchcggctggtccctgtggttctccgagtacctgttcctggagcgcaactgggccaaggacaagcgcaccctgaagtcccacatcgagcgcatgaaggactaccccctgcccctgtggctgatcctgttcgtggagggcacccgcttcacccgcaccaagctgctggccgcccagcagtacgccgcctcctccggcctgcccgtgccccgcaacgtgctgatcccccacaccaagggcttcgtgtcctccgtgtcccacatgcgctcatcgtgcccgccgtgtacgacgtgaccgtggcchccccaagacctcccccccccccaccatgctgtccctgttcgagggccagtccgtggtgctgcacgtgcacatcaagcgccacgccatgaaggacctgcccgactccgacgacgccgtggcccagtggtgccgcgacaagttcgtggagaaggacgccctgctggacaagcacaacgccgaggacaccttctccggccaggaggtgcaccacgtgggccgccccatcaagtccctgctggtggtgatctcctggatggtggtgatcatcttcggcgccctgaagttcctgcagtggtcctccctgctgtcctcctggaagggcaaggccttctccgccatcggcctgggcatcgccaccctgctgatgcacgtgctggtggtgttctcccaggccgaccgctccaaccccgccaaggtgccccccgccaagctgaacaccgagctgtcctcctccaagaaggtgaccaacaaggagaacTGA ctcgag CprocLPAAT2SEQ ID NO: 109 ggtaccATGgccatccccgccgccgtggccgtgatccccatcggcctgctgttcatcatctccggcctgatcgtgaacctgatccaggccgtggtgtacgtgctgatccgccccctgtccaagaacctgtaccgcaagatcaacaagcccatcgccgagctgctgtggctggagctgatctggctggtggactggtgggccggcatcaaggtggaggtgtacgccgactccgagaccctggagtccatgggcaaggagcacgccctgctgatctgcaaccaccgctccgacatcgactggctggtgggctgggtgctggcccagcgcgcccgctgcctgggctccgccctggccatcatgaagaagtccgccaagttcctgcccgtgatcggctggtccatgtggttctccgactacatcttcctggaccgcacctgggagaaggacgagaagaccctgaagtccggcttcgagcgcctggccgachccccatgcccttctggctggccctgttcgtggagggcacccgcttcaccaaggccaagctgctggccgcccaggagttcgccgcctcccgcggcctgcccgtgccccagaacgtgctgatcccccgcaccaagggcttcgtgaccgccgtgacccacatgcgctcctacgtgcccgccatctacgactgcaccgtggacatctccaaggcccaccccgccccctccatcctgcgcctgatccgcggccagtcctccgtggtgaaggtgcagatcacccgccactccatgcaggagctgcccgagacccccgacggcatctcccagtggtgcatggacctgttcgtgaccaaggacgcttcctggagaagtaccactccaaggacatcttcggctccctgcccgtgcacgacatcggccgccccgtgaagtccctgatcgtggtgctgtgctggtactccctgatggcchcggcactacaagttcttcatgtggtcctccctgctgtcctcctgggagggcatcctgtccctgggcctggtgctgatcgtgatcgccatcgtgatgcagatcctgatccagtcctccgagtccgagcgctccacccccgtgaagtccgtgcagaaggacccctccaaggagaccctgctgcagaacTGA ctcgag CavigGPAT9 SEQ ID NO: 110ggtaccATGgccaccggcggctccctgaagccctcctcctccgacctggacctggaccaccccaacatcgaggactacctgccctccggctcctccatcaacgagcccgccggcaagctgcgcctgcgcgacctgctggacatctcccccaccctgaccgaggccgccggcgccatcgtggacgactcchcacccgctgatcaagtccatcccccgcgagccctggaactggaacctgtacctgttccccctgtggtgcatcggcgtgctgatccgctacttcatcctgttccccggccgcgtgatcgtgctgaccatgggctggatcaccgtgatctcctcatcatcgccgtgcgcgtgctgctgaagggccacgacgccctgcagatcaagctggagcgcctgatcgtgcagctgctgtgctcctcatcgtggcctcctggaccggcgtggtgaagtaccacggcccccgcccctccatccgccccaagcaggtgtacgtggccaaccacacctccatgatcgacttchcatcctggaccagatgaccgtgttctccgtgatcatgcagaagcaccccggctgggtgggcctgctgcagtccaccctgctggagtccgtgggctgcatctggacgaccgcgccgaggccaaggaccgcggcatcgtggccaagaagctgtgggaccacgtgcacggcgagggcaacaaccccctgctgatchccccgagggcacctgcgtgaacaacaactactccgtgatgttcaagaagggcgcchcgagctgggctgcaccgtgtgccccgtggccatcaagtacaacaagatchcgtggacgcatctggaactccaagaagcagtcchcacccgccacctgctgcagctgatgacctcctgggccgtggtgtgcgacgtgtggtacttggagccccagaccctgaagcccggcgagacccccatcgagttcgccgagcgcgtgcgcgacatcatctccgcccgcgccggcctgaagaaggtgccctgggacggctacctgaagtactcccgcccctcccccaagcaccgcgagcgcaagcagcagacchcgccgagtccgtgctgcagcgcctggaggagTGA ctcgag ChookGPAT9-1 SEQ ID NO: 111 ggtaccATGgccaccgccggctccctgaagccctcccgctccgagctggacttcgaccgccccaacatcgaggactacctgccctccggctcctccatcatcgagcccgccggcaagctgcgcctgcgcgacctgctggacatctcccccaccctgaccgaggccgccggcgccatcgtggacgactcatcacccgctgatcaagtccaacccccccgagccctggaactggaacatctacctgttccccctgtggtgatcggcgtgctgatccgctacctgatcctgttccccgcccgcgtgatcgtgctgaccatcggctggatcatatcctgtcctcatcatccccgtgcacctgctgctgaagggccacgacgccctgcgcatcaagctggagcgcctgctggtggagctgatctgctcatatcgtggcctcctggaccggcgtggtgaagtaccacggcccccgcccctccatccgccccaagcaggtgtacgtggccaaccacacctccatgatcgacttatcatcctggaccagatgaccgtgttctccgtgatcatgcagaagcaccccggctgggtgggcctgctgcagtccaccctgctggagtccgtgggctgcatctggttcgaccgcgccgaggccaaggaccgcggcatcgtggccaagaagctgtgggaccacgtgcacggcgagggcaacaaccccctgctgatatccccgagggcacctgcgtgaacaacaactactccgtgatgttcaagaagggcgcatcgagctgggctgcaccgtgtgccccgtggccatcaagtacaacaagatatcgtggacgcatctggaactccaagaagcagtcatcacccgccacctgctgcagctgatgacctcctgggccgtggtgtgcgacgtgtggtacttggagccccagaccctgaagcccggcgagacccccatcgagttcgccgagcgcgtgcgcgacatcatctccgtgcgcgccggcctgaagaaggtgccctgggacggctacctgaagtactcccgcccctcccccaagcacaccgagcgcaagcagcagaacttcgccgagtccgtgctgcagcgcctggagaagaagTGA ctcgag CignGPAT9-1 SEQ ID NO: 112 ggtaccATGgccaccggcggccgcctgaagccctcctcctccgagctggacctggaccgcgccaacaccgaggactacctgccctccggctcctccatcaacgagcccgtgggcaagctgcgcctgcgcgacctgctggacatctcccccaccctgaccgaggccgccggcgccatcgtggacgactcatcacccgctgatcaagtccatcccccccgagccctggaactggaacatctacctgttccccctgtggtgatcggcgtgctgatccgctacttcatcctgttccccgcccgcgtgatcgtgctgaccatcggctggatcaccgtgatctcctcatcaccgccgtgcgatcctgctgaagggccacaacgccctgcagatcaagctggagcgcctgatcgtgcagctgctgtgctcctcatcgtggcctcctggaccggcgtggtgaagtaccacggcccccgcccctccatccgccccaagcaggtgtacgtggccaaccacacctccatgatcgacttcctgatcctggaccagatgaccgtgttctccgtgatcatgcagaagcaccccggctgggtgggcctgctgcagtccaccctgctggagtccgtgggctgcatctggttcaaccgcgccgaggccaaggaccgcgagatcgtggccaagaagctgtgggaccacgtgcacggcgagggcaacaaccccctgctgatatccccgagggcacctgcgtgaacaaccactactccgtgatgttcaagaagggcgcatcgagctgggctgcaccgtgtgccccgtggccatcaagtacaacaagatatcgtggacgcatctggaactcccgcaagcagtcatcaccatgcacctgctgcagctgatgacctcctgggccgtggtgtgcgacgtgtggtacttggagccccagaccctgaagcccggcgagaccgccatcgagttcgccgagcgcgtgcgcgacatcatctccgtgcgcgccggcctgaagaaggtgccctgggacggctacctgaagtactcccgcccctcccccaagcaccgcgagtccaagcagcagtcatcgccgagtccgtgctgcgccgcctggaggagaagTGA ctcgag CignGPAT9-2 SEQ ID NO: 113 ggtaccATGgccaccggcggccgcctgaagccctcctcctccgagctggacctggaccgcgccaacaccgaggactacctgccctccggctcctccatcaacgagcccgtgggcaagctgcgcctgcgcgacctgctggacatctcccccaccctgaccgaggccgccggcgccatcgtggacgactcatcacccgctgatcaagtccatcccccccgagccctggaactggaacatctacctgttccccctgtggtgatcggcgtgctgatccgctacttcatcctgttccccgcccgcgtgatcgtgctgaccatcggctggatcaccgtgatctcctcatcaccgccgtgcgatcctgctgaagggccacaacgccctgcagatcaagctggagcgcctgatcgtgcagctgctgtgctcctcatcgtggcctcctggaccggcgtggtgaagtaccacggcccccgcccctccatccgccccaagcaggtgtacgtggccaaccacacctccatgatcgacttcctgatcctggaccagatgaccgtgttctccgtgatcatgcagaagcaccccggctgggtgggcctgctgcagtccaccctgctggagtccgtgggctgcatctggttcaaccgcgccgaggccaaggaccgcgagatcgtggccaagaagctgtgggaccacgtgcacggcgagggcaacaaccccctgctgatatccccgagggcacctgcgtgaacaaccactactccgtgatgttcaagaagggcgcatcgagctgggctgcaccgtgtgccccgtggccatcaagtacaacaagatatcgtggacgcatctggaactccaagaagcactcatcacccgccacctgctgcagctgatgacctcctgggccgtggtgtgcgacgtgtggtacttggagccccagaccctgaagcccggcgagacccccatcgagttcgccgagcgcgtgcgcgacatcatctccgtgcgcgccgacctgaagaaggtgccctgggacggctacctgaagtactcccgcccctcccccaagcaccgcgagcgcaagcagcagaagttcgccgagtccgtgctgcgccgcctggaggagaagTGA ctcgag CpalGPAT9-1 SEQ ID NO: 114 ggtaccATGgccaccgccggccgcctgaagccctcctcctccgagctggagctggacctggaccgcccaacatcgaggactacctgccctccggctcctccatcaacgagcccgccggcaagctgcgcctgcgcgacctgctggacatctcccccatgctgaccgaggccgccggcgccatcgtggacgactccacacccgctgatcaagtccatcccccccgagccctggaactggaacatctacctgttccccctgtggtgatcggcgtgctgatccgctacctgatcctgaccccgcccgcgtgatcgtgctgaccgtgggctggatcaccgtgatctcctccacatcaccgtgcgcacctgctgaagggccacgactccctgcgcatcaagctggagcgcctgatcgtgcagctgttctgctcctccacgtggcctcctggaccggcgtggtgaagtaccacggcccccgcccctccatccgcccccagcaggtgtacgtggccaaccacacctccatgatcgacttcatcatcctgaaccagatgaccgtgactccgccatcatgcagaagcaccccggctgggtgggcctgatccagtccaccatcctggagtccgtgggctgcatctggacaaccgcgccgaggccaaggaccgcgagatcgtggccaagaagctgctggaccacgtgcacggcgagggcaacaaccccctgctgatcaccccgagggcacctgcgtgaacaaccactactccgtgatgacaagaagggcgccacgagctgggctgcaccgtgtgccccgtggccatcaagtacaacaagatcacgtggacgccttctggaactccaagaagcagtccacaccatgcacctgctgcagctgatgacctcctgggccgtggtgtgcgacgtgtggtacaggagccccagaccctgaagcccggcgagacccccatcgagacgccgagcgcgtgcgcgacatcatctccgtgcgcgccggcctgaagaaggtgccctgggacggctacctgaagtactcccgcccctcccccaagcaccgcgagcgcaagcagcagtccacgccgagtccgtgctgcgccgcctggagaagcgcTGA ctcgag CpalGPATt9-2 SEQ ID NO: 115ggtaccATGgccaccgccggccgcctgaagccctcctcctccgagctggagctggacctggaccgccccaacatcgaggactacctgccctccggctcctccatcaacgagcccgccggcaagctgcgcctgcgcgacctgctggacatctcccccatgctgaccgaggccgccggcgccatcgtggacgactccacacccgctgatcaagtccatcccccccgagccctggaactggaacatctacctgttccccctgtggtgatcggcgtgctgatccgctacctgatcctgaccccgcccgcgtgatcgtgctgaccgtgggctggatcaccgtgatctcctccacatcaccgtgcgcacctgctgaagggccacgactccctgcgcatcaagctggagcgcctgatcgtgcagctgttctgctcctccacgtggcctcctggaccggcgtggtgaagtaccacggcccccgcccctccatccgcccccagcaggtgtacgtggccaaccacacctccatgatcgacttcatcatcctgaaccagatgaccgtgactccgccatcatgcagaagcaccccggctgggtgggcctgatccagtccaccatcctggagtccgtgggctgcatctggacaaccgcgccgaggccaaggaccgcgagatcgtggccaagaagctgctggaccacgtgcacggcgagggcaacaaccccctgctgatcaccccgagggcacctgcgtgaacaaccactactccgtgatgacaagaagggcgccacgagctgggctgcaccgtgtgccccgtggccatcaagtacaacaagatcacgtggacgccttctggaactccaagaagctgtcatcaccatgcacctgctgcagctgatgacctcctgggccgtggtgtgcgacgtgtggtacaggagccccagaccctgaagcccggcgagacccccatcgagacgccgagcgcgtgcgcgacatcatctccgtgcgcgccggcctgaagaaggtgccctgggacggctacctgaagtactcccgcccctcccccaagcaccgcgagcgcaagcagcagaccacgccgagtccgtgctgcgccgcctggaggagaagggcaacgtggtgcccaccgtgaacTGA ctcgagCavigDGAT1 SEQ ID NO: 116 ggtaccATGgccatcgccgacggcggcatcatcggcgccgccggctccatctccgccctgaccgccgacaccgaccccccctccctgcgccgccgcaacgtgcccgccggccaggcctccgccgtgtccgccactccaccgagtccatggccaagcacctgtgcgacccctcccgcgagccctccccctcccccaagtcctccgacgacggcaaggaccccgacatcggctccgtggactccctgaacgagaagccctcctcccccgccgccggcaagggccgcctgcagcacgacctgcgcttcacctaccgcgcctcctcccccgcccaccgcaaggtgaaggagtcccccctgtcctcctccaacatcacaagcagtcccacgccggcctgacaacctgtgcgtggtggtgctggtggccgtgaactcccgcctgatcatcgagaacctgatgaagtacggcctgctgatcaagaccggataggactcctcccgctccctgcgcgactggcccctgacatgtgctgcctgtccctgcccatcaccccctggccgccacctggtggagaagctggcccagaagaaccgcctgcaggagcccaccgtggtgtgctgccacgtgctgatcacctccgtgtccatcctgtaccccgtgctggtgatcctgcgctgcgactccgccgtgctgtccggcgtggccctgatgctgacgcctgcatcgtgtggctgaagctggtgtcctacgcccactccaactacgacatgcgctacgtggccaagtccctggacaagggcgagcccgtggtggactccgtgatcgccgaccacccctaccgcgtggactacaaggacctggtgtacttcatggtggcccccaccctgtgctaccagctgtcctaccccctgaccccctgcgtgcgcaagtcctggatcgcccgccaggtgatgaagctggtgctgacaccggcgtgatgggatcatcgtggagcagtacatcaaccccatcgtgcagaactccaagcaccccctgaagggcgacctgctgtacgccatcgagcgcgtgctgaagctgtccgtgcccaacctgtacgtgtggctgtgcatgactactgatcaccacctgtggctgaacatcctggccgagctgatctgatcggcgaccgcgagactacaaggactggtggaacgccaagaccgtggaggagtactggcgcatgtggaacatgcccgtgcacaagtggatggtgcgccacatctacacccctgcctgcgcaacggcatcccccgcggcgtggccgtgctgatcgccacctggtgtccgccgtgaccacgagctgtgcatcgccgtgccctgccacgtgttcaagctgtgggcchcatcggcatcatgttccaggtgcccctggtgctggtgtccaactgcctgcagaagaagttccagtcctccatggccggcaacatgttatctgghcatchctgcatchcggccagcccatgtgcgtgctgctgtactaccacgacctgatgaaccgcaagggctccgcatcgacTGA ctcgag ChookDGAT1-1 SEQ ID NO: 117 ggtaccATGgccatcgccgacggcggctccgccggcgccgccggctccatctccggctccgacccctccccctccaccgccccctccctgcgccgccgcaacgcctccgccggccaggcchaccaccgagtccatggcccgcgacctgtgcgacccctcccgcgagccctccctgtcccccaagtcctccgacgacggcaaggaccccgccgacgacatcggcgccgccgactccgtggactccggcggcgtgaaggacgagaagccctcctcccaggccgccgccaaggcccgcctggagcacgacctgcgatcacctaccgcgcctcctcccccgcccaccgcaaggtgaaggagtcccccctgtcctcctccaacatchcaagcagtcccacgccggcctgttcaacctgtgcgtgggtggtgctggtggccgtgaactcccgcctgatcatcgagaacctgatgagtacggcctgctgataagaccggcttctggttctcctcccgctccctgcgcgactggcccctgttcatgtgctgcctgtccctgcccatcaccccctggccgcchcctggtggagaagctggcccagaagaaccgcctgcaggagcccaccgtggtgtgctgccacgtgatcatcacctccgtgtccatcctgtaccccgtgctggtgatcctgcgctgcgactccgccgtgctgtccggcgtggccctgatgctgttcgcctgcatcgtgtggctgaagctggtgtcctacgcccacgccaactacgacatgcgctccgtggccaagtccctggacaagggcgagaccgtggccgactccgtgatcgtggaccacccctaccgcgtggactacaaggacctggtgtacttcatggtggcccccaccctgtgctaccagctgtcctaccccctgaccccctacgtgcgcaagtcctgggtggcccgccaggtgatgaagctggtgctgttcaccggcgtgatgggchcatcgtggagcagtacatcaaccccatcgtgcagaactccaagcaccccctgaagggcgacctgctgtacgccatcgagcgcgtgctgaagctgtccgtgcccaacctgtacgtgtggctgtgcatgttctactgatchccacctgtggctgaacatcctggccgagctgacctgatcggcgaccgcgagttctacaaggactggtggaacgccaagaccgtggaggagtactggcgcatgtggaacatgcccgtgcacaagtggatggtgcgccacatctacttcccctgcctgcgcaacggcatcccccgcggcgtggccgtgctgatcgcchcctggtgtccgccgtgttccacgagctgtgcatcgccgtgccctgccacgtgttcaagctgtgggcchcatcggcatcatgttccaggtgcccctggtgctggtgtccaactgcctgcagaagaagttccagtcctccatggccggcaacatgttatctgghcatchctgcatchcggccagcccatgtgcgtgctgctgtactaccacgacctgatgaaccgaagggctcccgcatcgacTGA ctcgag CavigLPCATSEQ ID NO: 118ggtaccATGggcctggtgtccgtggccgccgccatcggcgtgtccgtgcccgtggcccgcttcctgctgtgcttcctggccaccatccccgtgtcchcctgtggcgcctggtgcccggccgcctgcccaagcacctgtactccgccgcctccggcgccatcctgtcctacctgtcatcggcgcctcctccaacctgcacttcatcgtgcccatgaccctgggctacctgtccatgctgttchccgcccatctccggcctgctgaccttcttcctgggcttcggctacctgatcggctgccacgtgtactacatgtccggcgacgcctggaaggagggcggcatcgacgccaccggcgccctgatggtgctgaccctgaaggtgatctcctgctccatgaactacaacgacggcctgctgaaggaggagggcctgcgcgagtcccagaagaagaaccgcctgaccaagatgccctccctgatcgagtacttcggctactgcctgtgctgcggctcccacttcgccggccccgtgtacgagatgaaggactacctggagtggaccgagggcaagggcatctggtcccgctcccagaaggagcccaagccctccccatcggcggcgccctgcgcgccatcatccaggccgccgtgtgcatggccatgtacctgtacctggtgccccaccaccccctgacccgatcaccgagcccgtgtactacgagtggggcttchccgccgcctgtcctaccagtacatggccgccctgaccgcccgctggaagtactacttcatctggtccatctccgaggcctccctgatcatctccggcctgggcttctccggctggaccgagtcctccccccccaagccccgctgggaccgcgccaagaacgtggacatcatcggcgtggagttcgccaagtcctccgtgcagctgcccctggtgtggaacatccaggtgtccatctggctgcgccactacgtgtacgaccgcctggtgcagaacggcaagcgccccggatcaccagctgctggccacccagaccgtgtccgccgtgtggcacggcctgtaccccggctacatcatchatcgtgcagtccgccctgatgatcgccggctcccgcgtgatctaccgctggcagcaggcggtgcccccaagatgggcctggtgaagaacatcttcgtgttcttcaacttcgcctacaccctgctggtgctgaactactccgccgtgggchcatggtgctgtccatgcacgagaccctggcctcctacggctccgtgtactacatcggcaccatcctgcccatcaccctgatcctgctgtcctacgtgatcaagcccggcaagcccgcccgctccaaggcccacaaggagcagTGA ctcgag CpalLPCAT SEQ ID NO: 119 ggtaccATGgagctgggctccgtggccgccgccatcggcgtgtccgtgcccgtggcccgcttcctgctgtgcttcctggccaccatccccgtgtcchcctgtggcgcctggtgcccggccgcctgcccaagcacctgtactccgccgcctccggcgccatcctgtcctacctgtccttcggcccctcctccaacctgcacttcatcgtgcccatgaccctgggctacctgtccatgctgttcttccgccccttctcggcctgctgaccttcttcctgggcttcggctacctgatcggctgccacgtgtactacatgtccggcgacgcctggaaggagggcggcatcgacgccaccggcgccctgatggtgctgaccctgaaggtgatctcctgctccatcaactacaacgacggcctgctgaaggaggagggcctgcgcgagtcccagaagaagaaccgcctgaccaagatgccctccctgatcgagtacatcggctactgcctgtgctgcggctcccacttcgccggccccgtgtacgagatgaaggactacctggagtggaccgagggcaagggcgtgtggtcccactccgagaaggagcccaagccctccccatcggcggcgccctgcgcgccatcatccaggccgccgtgtgcatggccatgtacatgtacctggtgccccaccaccccctgtcccgatcaccgagcccgtgtactacgagtggggcacttccgccgcctgtcctaccagtacatggccggcctgaccgcccgctggaagtactacttcatctggtccatctccgaggcctccctgatcatctccggcctgggcttctccggctggaccgagtcctccccccccaagccccgctgggaccgcgccaagaacgtggacatcatcggcgtggagttcgccaagtcctccgtgcagctgcccctggtgtggaacatccaggtgtccacctggctgcgccactacgtgtacgaccgcctggtgcagaacggcaagcgccccggatcaccagctgctggccacccagaccgtgtccgccatctggcacggcctgtaccccggctacatcatatatcgtgcagtccgccctgatgatcgccggctcccgcgtgatctaccgctggcagcaggccgtgccccccaagatgggcctggtgaagaacatcttcgtgttcttcaacttcgcctacaccctgctggtgctgaactactccgccgtgggatcatggtgctgtccatgcacgagaccctggcctcctacggctccgtgtactacatcggcaccatcctgcccatcaccctgatcctgctgtcctacgtgatcaagcccggcaagcccgcccgctccaaggcccacaaggagcagTGA ctcgag CpauLPCAT SEQ ID NO: 120 ggtaccATGgagctggagatcggctccgtggccgccgccatcggcgtgtccgtgcccgtggcccgcttcctgctgtgcttcctggccaccatccccgtgtccttcctgtgccgcctgctgcccgcccgcctgcccaagcacctgtactccgccgcctccggcgccatcctgtcctacctgtccttcggcccctcctccaacctgcacttcatcgtgcccatgtccctgggctacctgtccatgctgttcttccgccccttctccggcctgctgaccttcttcctgggcttcggctacctgatcggctgccacgtgtactacatgtccggcgacgcctggaaggagggcggcatcgacgccaccggcgccctgatggtgctgaccctgaaggtgatctcctgctccatcaactacaacgacggcctgctgaaggaggagggcctgcgcgagtcccagaagaagaaccgcctgaccaagatgccctccctgatcgagtacttcggctactgcctgtgctgcggctcccacttcgccggccccgtgtacgagatgaaggactacctggagtggaccgagggcaagggcatctggtcccgctccgagaaggaccccaagccctccccatcggcggcgccctgcgcgccatcatccaggccgccgtgtgcatggccatgcacatgtacctggtgccccaccaccccctgacccgcttcaccgagcccgtgtactacgagtggggcttcttccgccgcctgtcctaccagtacatggccgcccagaccgcccgctggaagtactacttcatctggtccatctccgaggcctccctgatcatctccggcctgggcttctccggctggaccgagtcctccccccccaagccccgctgggacaaggccaagaacgtggacatcatcggcgtggagttcgccaagtcctccgtgcagctgcccctggtgtggaacatccaggtgtccacctggctgcgccactacgtgtacgaccgcctggtgcagaacggcaagcgccccggcttcttccagctgctggccacccagaccgtgtccgccgtgtggcacggcctgtaccccggctacatcatcttcttcgtgcagtccgccctgatgatcgccggctcccgcgtgatctaccgctggcagcaggccgtgccccagaagatgggcctggtgaagaacatcttcgtgttcttcaacttcgcctacaccctgctggtgctgaactactccgccgtgggcttcatggtgctgtccatgcacgagaccctggcctcctacggctccgtgtactacatcggcaccatcctgcccatcaccctgatcctgctgtcctacgtgatcaagcccggcaagcccacccgctccaaggtgcacaaggagcagTGA ctcgag CschuLPCAT SEQ ID NO: 121 ggtaccATGgagctggagatggagcccctggccgccgccatcggcgtgtccgtggccgtgttccgcttcctggtgtgcttcatcgccaccatccccgtgtccttcatctgccgcctggtgcccggcggcctgccccgccacctgttctccgccgcctccggcgccgtgctgtcctacctgtccttcggcttctcctccaacctgcacttcctggtgcccatgaccctgggctacctgtccatgatcctgttccgccgcttctgcggcatcctgaccttcttcctgggcttcggctacctgatcggctgccacgtgtactacatgtccggcgacgcctggaaggagggcggcatcgacgccaccggcgccctgatggtgctgaccctgaaggtgatctcctgctccatcaactacaacgacggcctgctgaaggaggagggcctgcgcgagtcccagaagaagaaccgcctgatccgcctgccctccctgatcgagtacttcggctactgcctgtgctgcggctcccacttcgccggccccgtgtacgagatgaaggactacctggactggaccgagggcaagggcatctggtcccactccgagaagggccccaagccctcccccctgcgcgccgccctgcgcgccatcatccaggccggcttctgcatggccatgtacctgtacctggtgccccactaccccctgacccgcttcaccgaccccgtgtactacgagtggggcatcctgcgccgcctgtcctaccagtacatggcctccttcaccgcccgctggaagtactacttcatctggtccatctccgaggcctccctgatcatctccggcctgggcttctccggctggaccgagtcctccccccccaagccccgctgggaccgcgccaagaacgtggacatcctgggcgtggagctggccaagtcctccgtgcagatccccctggtgtggaacatccaggtgtccacctggctgcgccactacgtgtacgaccgcctggtgcagaacggcaagcgccccggatcctgcagctgctggccacccagaccgtgtccgccatctggcacggcgtgtaccccggctacctgatcacttcgtgcagtccgccctgatgatcgccggctcccgcgccatctaccgctggcagcaggccgtgccccccaagatgtccctggtgaagaacaccctggtgacttcaacttcgcctacaccctgctggtgctgaactactccgccgtgggatcatggtgctgtccatgcacgagaccctggcctcctacggctccgtgtactacgtgggcaccatcctgcccgtgaccctgatcctgctgggctacgtgatcaagcccggcaagtccccccgctccaaggcctccaaggagcagTGA ctcgag CavigPLA2-1 SEQ ID NO: 122 ggtaccATGaacttcgacttcctgtccaacatcccctggttcggcgccaaggcctccgacaacgccggctcctccttcggctccgccaccatcgtgatccagcagcccccccccgtgtcccgcggatcgacatccgccactggggctggccctggtccgtgctgtccgtgctgccctggggcaagcccggctgcgacgagctgcgcgccccccccaccaccatcaaccgccgcctgaagcgcaacgccacctccatgcactcctccgccgtgcgcggcaacgccgaggccgcccgcgtgcgcaccgcccctacgtgtccaaggtgccctggcacaccggcaccgcggcctgctgtcccagctgacccccgctacggccactactgcggccccaactggtcctccggcaagaacggcggctcccccgtgtgggaccagcgccccatcgactggctggactactgctgctactgccacgacatcggctacgacacccacgaccaggccaagctgctggaggccgacctggccacctggagtgcctggagcgcccctcctaccccaccaagggcgacgcccacgtggcccacatgtacaagaccatgtgcgtgaccggcctgcgcaacgtgctgatcccctaccgcacccagctgctgcgcctgaactcccgccagcccctgatcgacttcggctggctgtccaacgccgcctggaagggctggaacgcccagaagtccTGActcgag CignPLA2-1 SEQ ID NO: 123 ggtaccATGaacctggacttcctgtccaagatcccctggttcgaggccaaggcctccgagaaccccggcctgaacctgggctccaccaccatcgtgatcaagcagccccgccagggatcgacatccgccactggggctggccctggtccgtgctgacctggggcaaccgcgtgaccgacgaggtgcacgccccccccaccaccatcaaccgccgcctgaagcgcaacgccaccggccccgccgtgcagggcgacaccgaggccgcccgcctgcgcaccgcccctacgtgtccaaggtgccctggcacaccggcaccgcggcctgctgtcccagctgacccccgctacggccactactgcggccccaactggtcctccggcaagaacggcggctcccccgtgtgggaccagcgccccatcgactggctggactactgctgctactgccacgacatcggctacgacacccacgaccaggccaagctgctggaggccgacctggccacctggagtgcctggagcgcccctcctaccccaccaccggcgacgcccacgtggcccacatgtacaagaccatgtgcgtgaccggcctgcgcaacgtgctgatcccctaccgcacccagctgctgcgcctgaacttccgccagcccctgatcgacttcggctggctgtccaacgccgcctggaagggctggtccgcccagaagaccTGA ctcgag CuPSR23PLA2-2SEQ ID NO: 124 ggtaccATGgtgcacctgccccacaccctgaagctgggcctggtgatcgccatctccatctccggcctgtgcttctcctccacccccgcccgcgccctgaacgtgggcatccaggccgccggcgtgaccgtgtccgtgggcaagggctgctcccgcaagtgcgagtccgacactgcaaggtgccccccacctgcgctacggcaagtactgcggcctgatgtactccggctgccccggcgagaagccctgcgacggcctggacgcctgctgcatgaagcacgacgcctgcgtgcaggccaagaacaacgactacctgtcccaggagtgctcccagaacctgctgaactgcatggcctccaccgcatgtccggcggcaagcagacaagggctccacctgccaggtggacgaggtggtggacgtgctgaccgtggtgatggaggccgccctgctggccggccgctacctgcacaagcccTGA ctcgagCprocPLA2-2 SEQ ID NO: 125 ggtaccATGgtgcacctgccccacaccctgaagctgggcctggtgatcgccatctccatctccggcctgtgcctgtcctccacccccgcccgcgccctgaacgtgggcatccaggccgccggcgtgaccgtgtccgtgggcaagggctgctcccgcaagtgcgagtccgacactgcaaggtgccccccacctgcgctacggcaagtactgcggcctgatgtactccggctgccccggcgagaagccctgcgacggcctggacgcctgctgcatgaagcacgacgcctgcgtgcaggccaagaacgacgactacctgtcccaggagtgctcccagaacctgctgaactgcatggcctccaccgcatgtccggcggcaagcagacaagggctccacctgccaggtggacgaggtggtggacgtgctgaccgtggtgatggaggccgccctgctggccgccgctacctgcacaagcccTGA ctcgagNucleotide sequence of transforming DNA contained in pSZ5654 PmKASIISEQ ID NO: 126 gtttaaacgccggtcaccacccgcatgctcgtactacagcgcacgcaccgcttcgtgatccaccgggtgaacgtagtcctcgacggaaacatctggttcgggcctcctgcttgcactcccgcccatgccgacaacctttctgctgttaccacgacccacaatgcaacgcgacacgaccgtgtgggactgatcggttcactgcacctgcatgcaattgtcacaagcgcttactccaattgtattcgtttgttttctgggagcagttgctcgaccgcccgcgtcccgcaggcagcgatgacgtgtgcgtggcctgggtgtttcgtcgaaaggccagcaaccctaaatcgcaggcgatccggagattgggatctgatccgagtttggaccagatccgccccgatgcggcacgggaactgcatcgactcggcgcggaacccagctttcgtaaatgccagattggtgtccgatacctggatttgccatcagcgaaacaagacttcagcagcgagcgtatttggcgggcgtgctaccagggttgcatacattgcccatttctgtctggaccgctttactggcgcagagggtgagttgatggggttggcaggcatcgaaacgcgcgtgcatggtgtgcgtgtctgttttcggctgcacgaattcaatagtcggatgggcgacggtagaattgggtgtggcgctcgcgtgcatgcctcgccccgtcgggtgtcatgaccgggactggaatcccccctcgcgaccatcttgctaacgctcccgactc

gccactgcttcggcgcccgcctgcccaccgcctcccgccgcgccgtgcgccgcgcctggtcccgcatcgcccggggcgcgccaccgccgccgccgacgccaaccccgcccgccccgagcgccgcgtggtgatcaccggccagggcgtggtgacctccctgggccagaccatcgagcagttctactcctccctgctggagggcgtgtccggcatctcccagatccagaagttcgacaccaccggctacaccaccaccatcgccggcgagatcaagtccctgcagctggacccctacgtgcccaagcgctgggccaagcgcgtggacgacgtgatcaagtacgtgtacatcgccggcaagcaggccctggagtccgccggcctgcccatcgaggccgccggcctggccggcgccggcctggaccccgccctgtgcggcgtgctgatcggcaccgccatggccggcatgacctccttcgccgccggcgtggaggccctgacccgcggcggcgtgcgcaagatgaaccccttctgcatccccttctccatctccaacatgggcggcgccatgctggccatggacatcggcttcatgggccccaactactccatctccaccgcctgcgccaccggcaactactgcatcctgggcgccgccgaccacatccgccgcggcgacgccaacgtgatgctggccggcggcgccgacgccgccatcatcccctccggcatcggcggcttcatcgcctgcaaggccctgtccaagcgcaacgacgagcccgagcgcgcctcccgcccctgggacgccgaccgcgacggcttcgtgatgggcgagggcgccggcgtgctggtgctggaggagctggagcacgccaagcgccgcggcgccaccatcctggccgagctggtgggcggcgccgccacctccgacgcccaccacatgaccgagcccgacccccagggccgcggcgtgcgcctgtgcctggagcgcgccctggagcgcgcccgcctggcccccgagcgcgtgggctacgtgaacgcccacggcacctccacccccgccggcgacgtggccgagtaccgcgccatccgcgccgtgatcccccaggactccctgcgcatcaactccaccaagtccatgatcggccacctgctgggcggcgccggcgccgtggaggccgtggccgccatccaggccctgcgcaccggctggctgcaccccaacctgaacctggagaaccccgcccccggcgtggaccccgtggtgctggtgggcccccgcaaggagcgcgccgaggacctggacgtggtgctgtccaactccttcggcttcggcggccacaactcctgcgtgatcttccgcaagtacgacgagATGGACTACAAGGACCACGACGGCGACTACAA

cactctggacgctggtcgtgtgatggactgttgccgccacacttgctgccttgacctgtgaatatccctgccgcttttatcaaacagcctcagtgtgtttgatcttgtgtgtacgcgcttttgcgagttgctagctgcttgtgctatttgcgaataccacccccagcatccccttccctcgtttcatatcgcttgcatcccaaccgcaacttatctacgctgtcctgctatccctcagcgctgctcctgctcctgctcactgcccctcgcacagccttggtttgggctccgcctgtattctcctggtactgcaacctgtaaaccagcactgcaatgctgatgcacgggaagtagtgggatgggaacacaaatggagagctccgcgtctcgaacagagcgcgcagaggaacgctgaaggtctcgcctctgtcgcacctcagcgcggcatacaccacaataaccacctgacgaatgcgcttggttcttcgtccattagcgaagcgtccgttcacacacgtgccacgttggcg

cacgatcgggacgctgcgcaaggccatccccgcgcactgtttcgagcgctcggcgcttcgtagcagcatgtacctggcctttgacatcgcggtcatgtccctgctctacgtcgcgtcgacgtacatcgaccctgcaccggtgcctacgtgggtcaagtacggcatcatgtggccgctctactggttcttccaggtgtgtttgagggttttggttgcccgtattgaggtcctggtggcgcgcatggaggagaaggcgcctgtcccgctgacccccccggctaccctcccggcaccttccagggcgcgtacgagaagaaccagtagagcggccacatgatgccgtacttgacccacgtaggcaccggtgcagggtcgatgtacgtcgacgcgacgtagagcagggacatgaccgcgatgtcaaaggccaggtacatgctgctacgaagcgccgagcgctcgaaacagtgcgcggggatggccttgcgcagcgtcccgatcgtgaacggaggcttctccacaggctgcctgttcgtcttgatagccatctcgaggcagcagcagctcggatagtatcgacacactctggacgctggtcgtgtgatggactgttgccgccacacttgctgccttgacctgtgaatatccctgccgcttttatcaaacagcctcagtgtgtttgatcttgtgtgtacgcgcttttgcgagttgctagctgcttgtgctatttgcgaataccacccccagcatccccttccctcgtttcatatcgcttgcatcccaaccgcaacttatctacgctgtcctgctatccctcagcgctgctcctgctcctgctcactgcccctcgcacagccttggtttgggctccgcctgtattctcctggtactgcaacctgtaaaccagcactgcaatgctgatgcacgggaagtagtgggatgggaacacaaatggaaagctgtagagctcgatctaagtaagattcgaagcgctcgaccgtgccggacggactgcagccccatgtcgtagtgaccgccaatgtaagtgggctggcgtttccctgtacgtgagtcaacgtcactgcacgcgcaccaccctctcgaccggca

ctacaacggcctgggcctgacgccccagatgggctgggacaactggaacacgttcgcctgcgacgtctccgagcagctgctgctggacacggccgaccgcatctccgacctgggcctgaaggacatgggctacaagtacatcatcctggacgactgctggtcctccggccgcgactccgacggcttcctggtcgccgacgagcagaagttccccaacggcatgggccacgtcgccgaccacctgcacaacaactccttcctgttcggcatgtactcctccgcgggcgagtacacgtgcgccggctaccccggctccctgggccgcgaggaggaggacgcccagttcttcgcgaacaaccgcgtggactacctgaagtacgacaactgctacaacaagggccagttcggcacgcccgagatctcctaccaccgctacaaggccatgtccgacgccctgaacaagacgggccgccccatcttctactccctgtgcaactggggccaggacctgaccttctactggggctccggcatcgcgaactcctggcgcatgtccggcgacgtcacggcggagttcacgcgccccgactcccgctgcccctgcgacggcgacgagtacgactgcaagtacgccggcttccactgctccatcatgaacatcctgaacaaggccgcccccatgggccagaacgcgggcgtcggcggctggaacgacctggacaacctggaggtcggcgtcggcaacctgacggacgacgaggagaaggcgcacttctccatgtgggccatggtgaagtcccccctgatcatcggcgcgaacgtgaacaacctgaaggcctcctcctactccatctactcccaggcgtccgtcatcgccatcaaccaggactccaacggcatccccgccacgcgcgtctggcgctactacgtgtccgacacggacgagtacggccagggcgagatccagatgtggtccggccccctggacaacggcgaccaggtcgtggcgctgctgaacggcggctccgtgtcccgccccatgaacacgaccctggaggagatcttcttcgactccaacctgggctccaagaagctgacctccacctgggacatctacgacctgtgggcgaaccgcgtcgacaactccacggcgtccgccatcctgggccgcaacaagaccgccaccggcatcctgtacaacgccaccgagcagtcctacaaggacggcctgtccaagaacgacacccgcctgttcggccagaagatcggctccctgtcccccaacgcgatcctgaacacgaccgtccccgcccacggcatcgcgttct

ttactcttgaggaattgaacctttctcgcttgctggcatgtaaacattggcgcaattaattgtgtgatgaagaaagggtggcacaagatggatcgcgaatgtacgagatcgacaacgatggtgattgttatgaggggccaaacctggctcaatcttgtcgcatgtccggcgcaatgtgatccagcggcgtgactctcgcaacctggtagtgtgtgcgcaccgggtcgctttgattaaaactgatcgcattgccatcccgtcaactcacaagcctactctagctcccattgcgcactcgggcgcccggctcgatcaatgttctgagcggagggcgaagcgtcaggaaatcgtctcggcagctggaagcgcatggaatgcggagcggagatcgaatcaggatccttagggagcgacgagtgtgcgtgcggggctggcgggagtgggacgccctcctcgctcctctctgttctgaacggaacaatcggccaccccgcgctacgcgccacgcatcgagcaacgaagaaaaccccccgatgataggttgcggtggctgccgggatatagatccggccgcacatcaaagggcccctccgccagagaagaagctcctttcccagcagactccttctgctgccaaaacacttctctgtccacagcaacaccaaaggatgaacagatcaacttgcgtctccgcgtagcttcctcggctagcgtgcttgcaacaggtccctgcactattatcttcctgctttcctctgaattatgcggcaggcgagcgctcgctctggcgagcgctccttcgcgccgccctcgctgatcgagtgtacagtcaatgaatggtcctgggcgaagaacgagggaatttgtgggtaaaacaagcatcgtctctcaggccccggcgcagtggccgttaaagtccaagaccgtgaccaggcagcgcagcgcgtccgtgtgcgggccctgcctggcggctcggcgtgccaggctcgagagcagctccctcaggtcgccttggacggcctctgcgaggccggtgagggcctgcaggagcgcctcgagcgtggcagtggcggtcgtatccgggtcgccggtcaccgcctgcgactcgccatccgaagagcgtttaaac Nucleotide sequence of transforming DNA contained in pSZ5868GarmFATA1(G108A) SEQ ID NO: 127 gaagagc gcccaat gtttaaacctcttttgctgcgtctcctcaggcttgggggcctccttgggcttgggtgccgccatgatctgcgcgcatcagagaaacgttgctggtaaaaaggagcgcccggctgcgcaatatatatataggcatgccaacacagcccaacctcactcgggagcccgtcccaccacccccaagtcgcgtgccttgacggcatactgctgcagaagcttcatgagaatgatgccgaacaagaggggcacgaggacccaatcccggacatccttgtcgataatgatctcgtgagtccccatcgtccgcccgacgctccggggagcccgccgatgctcaagacgagagggccctcgaccaggaggggctggcccgggcgggcactggcgtcgaaggtgcgcccgtcgttcgcctgcagtcctatgccacaaaacaagtcttctgacggggtgcgtttgctcccgtgcgggcaggcaacagaggtattcaccctggtcatggggagatcggcgatcgagctgggataagagatacggtcccgcgcaaggatcgctcatcctggtctgagccggacagtcattctggcaagcaatgacaacttgtcaggaccggaccgtgccatatatttctcacctagcgccgcaaaacctaacaatttgggagtcactgtgccactgagttcgactggtagctgaatggagtcgctgctccactaaacgaattgtcagcaccgccagccggccgaggacccgagtcata

ggcgggctccgggccccggcgcccagcgaggcccctccccgtgcgcgggcgcgccatccccccccgcatcatcgtggtgtcctcctcctcctccaaggtgaaccccctgaagaccgaggccgtggtgtcctccggcctggccgaccgcctgcgcctgggctccctgaccgaggacggcctgtcctacaaggagaagttcatcgtgcgctgctacgaggtgggcatcaacaagaccgccaccgtggagaccatcgccaacctgctgcaggaggtgggctgcaaccacgcccagtccgtgggctactccaccgccggcttctccaccacccccaccatgcgcaagctgcgcctgatctgggtgaccgcccgcatgcacatcgagatctacaagtaccccgcctggtccgacgtggtggagatcgagtcctggggccagggcgagggcaagatcggcacccgccgcgactggatcctgcgcgactacgccaccggccaggtgatcggccgcgccacctccaagtgggtgatgatgaaccaggacacccgccgcctgcagaaggtggacgtggacgtgcgcgacgagtacctggtgcactgcccccgcgagctgcgcctggccttccccgaggagaacaactcctccctgaagaagatctccaagctggaggacccctcccagtactccaagctgggcctggtgccccgccgcgccgacctggacatgaaccagcacgtgaacaacgtgacctacatcggctgggtgctggagtccatgccccaggagatcatcgacacccacgagctgcagaccatcaccctggactaccgccgcgagtgccagcacgacgacgtggtggactccctgacctcccccgagccctccgaggacgccgaggccgtgttcaaccacaacggcaccaacggctccgccaacgtgtccgccaacgaccacggctgccgcaacttcctgcacctgctgcgcctgtccggcaacggcctggagatcaaccgcggccgcaccgagtggcgcaagaagcccacccgcATGGACTACAAGGACCACGACGGCGACTACAAGGACCACGACATCGACTACAAGGACGACGACGACAAG TGAatcgatggagcgacgagtgtgcgtgcggggctggcgggagtgggacgccctcctcgctcctctctgttctgaacggaacaatcggccaccccgcgctacgcgccacgcatcgagcaacgaagaaaaccccccgatgataggttgcggtggctgccgggatatagatccggccgcacatcaaagggcccctccgccagagaagaagctcctttcccagcagactccttctgctgccaaaacacttctctgtccacagcaacaccaaaggatgaacagatcaacttgcgtctccgcgtagcttcctcggctagcgtgcttgcaacaggtccctgcactattatcttcctgctttcctctgaattatgcggcaggcgagcgctcgctctggcgagcgctccttcgcgccgccctcgctgatcgagtgtacagtcaatgaatggtgagctccgcgtctcgaacagagcgcgcagaggaacgctgaaggtctcgcctctgtcgcacctcagcgcggcatacaccacaataaccacctgacgaatgcgcttggttcttcgtccattagcgaagcgtccggttcacacacgtgccacgttggcgaggtggcaggtgacaatgatcggtggagctgatggt

tcgccgccgccgccgtgatcgtgcccctgggcctgctgttcttcatctccggcctggtggtgaacctgatccaggccctgtgcttcgtgctgatccgccccctgtccaagaacacctaccgcaagatcaaccgcgtggtggccgagctgctgtggctggagctgatctggctggtggactggtgggccggcgtgaagatcaaggtgttcatggaccccgagtccttcaacctgatgggcaaggagcacgccctggtggtggccaaccaccgctccgacatcgactggctggtgggctggctgctggcccagcgctccggctgcctgggctccgccctggccgtgatgaagaagtcctccaagttcctgcccgtgatcggctggtccatgtggttctccgagtacctgttcctggagcgctcctgggccaaggacgagaacaccctgaaggccggcctgcagcgcctgaaggacttcccccgccccttctggctggccttcttcgtggagggcacccgcttcacccaggccaagttcctggccgcccaggagtacgccgcctcccagggcctgcccatcccccgcaacgtgctgatcccccgcaccaagggcttcgtgtccgccgtgtcccacatgcgctccttcgtgcccgccatctacgacatgaccgtggccatccccaagtcctccccctcccccaccatgctgcgcctgttcaagggccagccctccgtggtgcacgtgcacatcaagcgctgcctgatgaaggagctgcccgagaccgacgaggccgtggcccagtggtgcaaggacatgacgtggagaaggacaagctgctggacaagcacatcgccgaggacaccactccgaccagcccatgcaggacctgggccgccccatcaagtccctgctggtggtggcctcctgggcctgcctgatggcctacggcgccctgaagttcctgcagtgctcctccctgctgtcctcctggaagggcatcgccacttcctggtgggcctggccatcgtgaccatcctgatgcacatcctgatcctgttctcccagtccgagcgctccacccccgccaaggtgg

agggtggtcgactcgttggaggtgggtgtttttttttatcgagtgcgcggcgcggcaaacgggtccctttttatcgaggtgttcccaacgccgcaccgccctcttaaaacaacccccaccaccacttgtcgaccttctcgtttgttatccgccacggcgccccggaggggcgtcgtctggccgcgcgggcagctgtatcgccgcgctcgctccaatggtgtgtaatcttggaaagataataatcgatggatgaggaggagagcgtgggagatcagagcaaggaatatacagttggcacgaagcagcagcgtactaagctgtagcgtgttaagaaagaaaaactcgctgttaggctgttaatcaaggagcgtatcaataattaccgaccctatacctttatctccaacccaatcgcggcttaaggatctaagtaagattcgaagcgctcgaccgtgccggacggactgcagccccatgtcgtagtgaccgccaatgtaagtgggctggcgtttccctgtacgtgagtcaacgtcactgcacgcgcaccaccctctcgaccggcaggaccaggcatcgcgagatacagcgcgagccagacacggagtg

ccgaccgccccctggtgcacttcacccccaacaagggctggatgaacgaccccaacggcctgtggtacgacgagaaggacgccaagtggcacctgtacttccagtacaacccgaacgacaccgtctgggggacgcccttgactggggccacgccacgtccgacgacctgaccaactgggaggaccagcccatcgccatcgccccgaagcgcaacgactccggcgccttctccggctccatggtggtggactacaacaacacctccggcttcttcaacgacaccatcgacccgcgccagcgctgcgtggccatctggacctacaacaccccggagtccgaggagcagtacatctcctacagcctggacggcggctacaccttcaccgagtaccagaagaaccccgtgctggccgccaactccacccagttccgcgacccgaaggtcttctggtacgagccctcccagaagtggatcatgaccgcggccaagtcccaggactacaagatcgagatctactcctccgacgacctgaagtcctggaagctggagtccgcgttcgccaacgagggcacctcggctaccagtacgagtgccccggcctgatcgaggtccccaccgagcaggaccccagcaagtcctactgggtgatgttcatctccatcaaccccggcgccccggccggcggctccttcaaccagtacttcgtcggcagcttcaacggcacccacttcgaggccttcgacaaccagtcccgcgtggtggacttcggcaaggactactacgccctgcagaccttcttcaacaccgacccgacctacgggagcgccctgggcatcgcgtgggcctccaactgggagtactccgccacgtgcccaccaacccctggcgctcctccatgtccctcgtgcgcaagactccctcaacaccgagtaccaggccaacccggagacggagctgatcaacctgaaggccgagccgatcctgaacatcagcaacgccggcccctggagccggttcgccaccaacaccacgttgacgaaggccaacagctacaacgtcgacctgtccaacagcaccggcaccctggagttcgagctggtgtacgccgtcaacaccacccagacgatctccaagtccgtgttcgcggacctctccctctggttcaagggcctggaggaccccgaggagtacctccgcatgggcttcgaggtgtccgcgtcctccttcttcctggaccgcgggaacagcaaggtgaagttcgtgaaggagaacccctacttcaccaaccgcatgagcgtgaacaaccagcccttcaagagcgagaacgacctgtcctactacaaggtgtacggcttgctggaccagaacatcctggagctgtacttcaacgacggcgacgtcgtgtccaccaacacctacttcatgaccaccgggaacgccctgggctccgtgaacatgacgacgggggtggacaacctgttctacatcgaca

cgaaacaagcccctggagcatgcgtgcatgatcgtctctggcgccccgccgcgcggtttgtcgccctcgcgggcgccgcggccgcgggggcgcattgaaattgttgcaaaccccacctgacagattgagggcccaggcaggaaggcgttgagatggaggtacaggagtcaagtaactgaaagtttttatgataactaacaacaaagggtcgtttctggccagcgaatgacaagaacaagattccacatttccgtgtagaggcttgccatcgaatgtgagcgggcgggccgcggacccgacaaaacccttacgacgtggtaagaaaaacgtggcgggcactgtccctgtagcctgaagaccagcaggagacgatcggaagcatcacagcacaggatcctgaggacagggtggttggctggatggggaaacgctggtcgcgggattcgatcctgctgcttatatcctccctggaagcacacccacgactctgaagaagaaaacgtgcacacacacaacccaaccggccgaatatttgcttccttatcccgggtccaagagagactgcgatgcccccctcaatcagcatcctcctccctgccgcttcaatcttccctgcttgcctgcgcccgcggtgcgccgtctgcccgcccagtcagtcactcctgcacaggccccttgtgcgcagtgctcctgtaccctttaccgctccttccattctgcgaggccccctattgaatgtattcgttgcctgtgtggccaagcgggctgctgggcgcgccgccgtcgggcagtgctcggcgactttggcggaagccgattgttcttctgtaagccacgcgcttgctgctttgggaagagaagggggggggtactgaatggatgaggaggagaaggaggggtattggtattatctgagttggggaggcagggagagttggaaaatgtaagtggcacgacgggcaaggagaatggtgagcatgtgcatggtgatgtcgttggtcgaggacgatcctgcacgcgtgtatctgatgtagaatacggcaatcaccctagtctacatctataccttctccgtataacgccctttccaaatgccctcccgtttctctcctattcttgatccacatgatgaccctggcactatttcaagggctgga gaagagcgtttaaacNucleotide sequence of transforming DNA contained in pSZ6383TcDGAT1 and GarmFATA1(G108A) SEQ ID NO: 128 gctcttcgcgaaggtcattttccagaacaacgaccatggcttgtcttagcgatcgctcgaatgactgctagtgagtcgtacgctcgacccagtcgctcgcaggagaacgcggcaactgccgagcttcggcttgccagtcgtgactcgtatgtgatcaggaatcattggcattggtagcattataattcggcttccgcgctgtttatgggcatggcaatgtctcatgcagtcgaccttagtcaaccaattctgggtggccagctccgggcgaccgggctccgtgtcgccgggcaccacctcctgccatgagtaacagggccgccctctcctcccgacgttggccaactgaataccgtgtcttggggccctacatgatgggctgcctagtcgggcgggacgcgcaactgcccgcgcaatctgggacgtggtctgaatcctccaggcgggtttccccgagaaagaaagggtgccgatttcaaagcagagccatgtgccgggccctgtggcctgtgttggcgcctatgtagtcaccccccctcacccaattgtcgccagtttgcgcaatccataaactcaaaactgcagcttctgagctgcgctgttcaa

ctgatgtccgtggtctgcaacaacaagaaccactccgcccgccccaagctgcccaactcctccctgctgcccggcttcgacgtggtggtccaggccgcggccacccgcttcaagaaggagacgacgaccacccgcgccacgctgacgttcgacccccccacgaccaactccgagcgcgccaagcagcgcaagcacaccatcgacccctcctcccccgacttccagcccatcccctccttcgaggagtgcttccccaagtccacgaaggagcacaaggaggtggtgcacgaggagtccggccacgtcctgaaggtgcccttccgccgcgtgcacctgtccggcggcgagcccgccttcgacaactacgacacgtccggcccccagaacgtcaacgcccacatcggcctggcgaagctgcgcaaggagtggatcgaccgccgcgagaagctgggcacgccccgctacacgcagatgtactacgcgaagcagggcatcatcacggaggagatgctgtactgcgcgacgcgcgagaagctggaccccgagttcgtccgctccgaggtcgcgcggggccgcgccatcatcccctccaacaagaagcacctggagctggagcccatgatcgtgggccgcaagttcctggtgaaggtgaacgcgaacatcggcaactccgccgtggcctcctccatcgaggaggaggtctacaaggtgcagtgggccaccatgtggggcgccgacaccatcatggacctgtccacgggccgccacatccacgagacgcgcgagtggatcctgcgcaactccgcggtccccgtgggcaccgtccccatctaccaggcgctggagaaggtggacggcatcgcggagaacctgaactgggaggtgttccgcgagacgctgatcgagcaggccgagcagggcgtggactacttcacgatccacgcgggcgtgctgctgcgctacatccccctgaccgccaagcgcctgacgggcatcgtgtcccgcggcggctccatccacgcgaagtggtgcctggcctaccacaaggagaacttcgcctacgagcactgggacgacatcctggacatctgcaaccagtacgacgtcgccctgtccatcggcgacggcctgcgccccggctccatctacgacgccaacgacacggcccagttcgccgagctgctgacccagggcgagctgacgcgccgcgcgtgggagaaggacgtgcaggtgatgaacgagggccccggccacgtgcccatgcacaagatccccgagaacatgcagaagcagctggagtggtgcaacgaggcgcccttctacaccctgggccccctgacgaccgacatcgcgcccggctacgaccacatcacctccgccatcggcgcggccaacatcggcgccctgggcaccgccctgctgtgctacgtgacgcccaaggagcacctgggcctgcccaaccgcgacgacgtgaaggcgggcgtcatcgcctacaagatcgccgcccacgcggccgacctggccaagcagcacccccacgcccaggcgtgggacgacgcgctgtccaaggcgcgcttcgagttccgctggatggaccagttcgcgctgtccctggaccccatgacggcgatgtccttccacgacgagacgctgcccgcggacggcgcgaaggtcgcccacttctgctccatgtgcggccccaagttctgctccatgaagatcacggaggacatccgcaagtacgccgaggagaacggctacggctccgccgaggaggccatccgccagggcatggacgccatgtccgaggagttcaacatcgccaagaagacgatctccggcgagcagcacggcgaggtcggcggcgagatctacctgcccgagtcctacgtc

cgcacgcatccaacgaccgtatacgcatcgtccaatgaccgtcggtgtcctctctgcctccgttttgtgagatgtctcaggcttggtgcatcctcgggtggccagccacgttgcgcgtcgtgctgcttgcctctcttgcgcctctgtggtactggaaaatatcatcgaggcccgtttttttgctcccatttcctttccgctacatcttgaaagcaaacgacaaacgaagcagcaagcaaagagcacgaggacggtgaacaagtctgtcacctgtatacatctatttccccgcgggtgcacctactctctcctgccccggcagagtcagctgccttacgtgacggatcccgcgtctcgaacagagcgcgcagaggaacgctgaaggtctcgcctctgtcgcacctcagcgcggcatacaccacaataaccacctgacgaatgcgcttggttcttcgtccattagcgaagcgtccggttcacacacgtgccacgttggcgaggtggcaggtgacaatgatcggtgga

cgagatcctgggctccaccgccaccgtgacctcctcctcccactccgactccgacctgaacctgctgtccatccgccgccgcacctccaccaccgccgccgcccgcgcccccgaccgcgacgactccggcaacggcgaggccgtggacgaccgcgaccgcgtggagtccgccaacctgatgtccaacgtggccgagaacgccaacgagatgcccaactcctccgacacccgcttcacctaccgcccccgcgtgcccgcccaccgccgcatcaaggagtcccccctgtcctccggcgccatcttcaagcagtcccacgccggcctgttcaacctgtgcatcgtggtgctggtggccgtgaactcccgcctgatcatcgagaacctgatgaagtacggctggctgatccgctccggcttctggttctcctcccgctccctgtccgactggcccctgttcatgtgctgcctgaccctgcccatcttccccctggccgccttcgtggtggagaagctggtgcagcgcaactacatctccgagcccgtggtggtgttcctgcacgccatcatctccaccaccgccgtgctgtaccccgtgatcgtgaacctgcgctgcgactccgccttcctgtccggcgtggccctgatgctgttcgcctgcatcgtgtggctgaagctggtgtcctacgcccacaccaacaacgacatgcgcgccctggccaagtccgccgagaagggcgacgtggacccctcctacgacgtgtccttcaagtccctggcctacttcatggtggcccccaccctgtgctaccagcagtcctacccccgcacccccgccgtgcgcaagtcctgggtggtgcgccagttcatcaagctgatcgtgttcaccggcctgatgggcttcatcatcgagcagtacatcaaccccatcgtgcagaactcccagcaccccctgaagggcaacctgctgtacgccatcgagcgcgtgctgaagctgtccgtgcccaacctgtacgtgtggctgtgcatgttctactgcttcttccacctgtggctgaacatcctggccgagctgctgcgcttcggcgaccgcgagttctacaaggactggtggaacgccaagaccgtggaggagtactggcgcatgtggaacatgcccgtgcacaagtggatggtgcgccacatctacttcccctgcctgcgcaacggcatccccaagggcgtggccatcgtgatcgccttcctggtgtccgccgtgttccacgagctgtgcatcgccgtgccctgccacatgttcaagctgtgggccttcatcggcatcatgttccaggtgcccctggtgctgatcaccaactacctgcaggacaagttccgctcctccatggtgggcaacatgatcttctggttcatcttctccatcctgggccagcccatgtgcgtgctgctgt

gacacactctggacgctggtcgtgtgatggactgttgccgccacacttgctgccttgacctgtgaatatccctgccgcttttatcaaacagcctcagtgtgtttgatcttgtgtgtacgcgcttttgcgagttgctagctgcttgtgctatttgcgaataccacccccagcatccccttccctcgtttcatatcgcttgcatcccaaccgcaacttatctacgctgtcctgctatccctcagcgctgctcctgctcctgctcactgcccctcgcacagccttggtttgggctccgcctgtattctcctggtactgcaacctgtaaaccagcactgcaatgctgatgcacgggaagtagtgggatgggaacacaaatggacttaaggatctaagtaagattcgaagcgctcgaccgtgccggacggactgcagccccatgtcgtagtgaccgccaatgtaagtgggctggcgtttccctgtacgtgagtcaacgtcactgcacgcgcaccaccctctcgaccggcaggaccaggcatcgcgagatacagcgcgagccagacacggagtgccgagctatgcgcacgctccaactagatatcatgtggatgatgagcat

aatgcccgctgcggcgacctgcgtcgctcggcgggctccagggccccggcgcccagcgaggcccctccccgtgcgcggcgcgccatccccccccgcatcatcgtggtgtcctcctcctcctccaaggtgaaccccctgaagaccgaggccgtggtgtcctccggcctggccgaccgcctgcgcctgggctccctgaccgaggacggcctgtcctacaaggagaagttcatcgtgcgctgctacgaggtgggcatcaacaagaccgccaccgtggagaccatcgccaacctgctgcaggaggtgggctgcaaccacgcccagtccgtgggctactccaccgccggcttctccaccacccccaccatgcgcaagctgcgcctgatctgggtgaccgcccgcatgcacatcgagatctacaagtaccccgcctggtccgacgtggtggagatcgagtcctggggccagggcgagggcaagatcggcacccgccgcgactggatcctgcgcgactacgccaccggccaggtgatcggccgcgccacctccaagtgggtgatgatgaaccaggacacccgccgcctgcagaaggtggacgtggacgtgcgcgacgagtacctggtgcactgcccccgcgagctgcgcctggccttccccgaggagaacaactcctccctgaagaagatctccaagctggaggacccctcccagtactccaagctgggcctggtgccccgccgcgccgacctggacatgaaccagcacgtgaacaacgtgacctacatcggctgggtgctggagtccatgccccaggagatcatcgacacccacgagctgcagaccatcaccctggactaccgccgcgagtgccagcacgacgacgtggtggactccctgacctcccccgagccctccgaggacgccgaggccgtgttcaaccacaacggcaccaacggctccgccaacgtgtccgccaacgaccacggctgccgcaacttcctgcacctgctgcgcctgtccggcaacggcctggagatcaaccgcggccgcaccgagtggcgcaagaagcccacccgcATGGACTACAAGGACCACGACGGCGACTACAAGGACCACGACATCGACTACAAGGACGACGACGACAA

tcggccaccccgcgctacgcgccacgcatcgagcaacgaagaaaaccccccgatgataggttgcggtggctgccgggatatagatccggccgcacatcaaagggcccctccgccagagaagaagctcctttcccagcagactccttctgctgccaaaacacttctctgtccacagcaacaccaaaggatgaacagatcaacttgcgtctccgcgtagcttcctcggctagcgtgcttgcaacaggtccctgcactattatcttcctgctttcctctgaattatgcggcaggcgagcgctcgctctggcgagcgctccttcgcgccgccctcgctgatcgagtgtacagtcaatgaatggtgagctcctcactcagcgcgcctgcgcggggatgcggaacgccgccgccgccttgtcttttgcacgcgcgactccgtcgcttcgcgggtggcacccccattgaaaaaaacctcaattctgtttgtggaagacacggtgtacccccaaccacccacctgcacctctattattggtattattgacgcgggagcgggcgttgtactctacaacgtagcgtctctggttttcagctggctcccaccattgtaaattcttgctaaaatagtgcgtggttatgtgagaggtatggtgtaacagggcgtcagtcatgttggttttcgtgctgatctcgggcacaaggcgtcgtcgacgtgacgtgcccgtgatgagagcaataccgcgctcaaagccgacgcatggcctttactccgcactccaaacgactgtcgctcgtatttttcggatatctattttttaagagcgagcacagcgccgggcatgggcctgaaaggcctcgcggccgtgctcgtggtgggggccgcgagcgcgtggggcatcgcggcagtgcaccaggcgcagacggaggaacgcatggtgagtgcgcatcacaagatgcatgtcttgttgtctgtactataatgctagagcatcaccaggggcttagtcatcgcacctgctttggtcattacagaaattgcacaagggcgtcctccgggatgaggagatgtaccagctcaagctggagcggcttcgagccaagcaggagcgcggcgcatgacgacctacccacatgc gaagagcNucleotide sequence of transforming DNA contained in pSZ6384TcDGAT2-and GarmFATA1(G108A) SEQ ID NO: 129 gctcttcgcgaaggtcattttccagaacaacgaccatggcttgtcttagcgatcgctcgaatgactgctagtgagtcgtacgctcgacccagtcgctcgcaggagaacgcggcaactgccgagcttcggcttgccagtcgtgactcgtatgtgatcaggaatcattggcattggtagcattataattcggcttccgcgctgtttatgggcatggcaatgtctcatgcagtcgaccttagtcaaccaattctgggtggccagctccgggcgaccgggctccgtgtcgccgggcaccacctcctgccatgagtaacagggccgccctctcctcccgacgttggccaactgaataccgtgtcttggggccctacatgatgggctgcctagtcgggcgggacgcgcaactgcccgcgcaatctgggacgtggtctgaatcctccaggcgggtttccccgagaaagaaagggtgccgatttcaaagcagagccatgtgccgggccctgtggcctgtgttggcgcctatgtagtcaccccccctcacccaattgtcgccagtttgcgcaatccataaactcaaaactgcagcttctgagctgcgctgttcaa

ctgatgtccgtggtctgcaacaacaagaaccactccgcccgccccaagctgcccaactcctccctgctgcccggcttcgacgtggtggtccaggccgcggccacccgcttcaagaaggagacgacgaccacccgcgccacgctgacgttcgacccccccacgaccaactccgagcgcgccaagcagcgcaagcacaccatcgacccctcctcccccgacttccagcccatcccctccttcgaggagtgcttccccaagtccacgaaggagcacaaggaggtggtgcacgaggagtccggccacgtcctgaaggtgcccttccgccgcgtgcacctgtccggcggcgagcccgccttcgacaactacgacacgtccggcccccagaacgtcaacgcccacatcggcctggcgaagctgcgcaaggagtggatcgaccgccgcgagaagctgggcacgccccgctacacgcagatgtactacgcgaagcagggcatcatcacggaggagatgctgtactgcgcgacgcgcgagaagctggaccccgagttcgtccgctccgaggtcgcgcggggccgcgccatcatcccctccaacaagaagcacctggagctggagcccatgatcgtgggccgcaagttcctggtgaaggtgaacgcgaacatcggcaactccgccgtggcctcctccatcgaggaggaggtctacaaggtgcagtgggccaccatgtggggcgccgacaccatcatggacctgtccacgggccgccacatccacgagacgcgcgagtggatcctgcgcaactccgcggtccccgtgggcaccgtccccatctaccaggcgctggagaaggtggacggcatcgcggagaacctgaactgggaggtgttccgcgagacgctgatcgagcaggccgagcagggcgtggactacttcacgatccacgcgggcgtgctgctgcgctacatccccctgaccgccaagcgcctgacgggcatcgtgtcccgcggcggctccatccacgcgaagtggtgcctggcctaccacaaggagaacttcgcctacgagcactgggacgacatcctggacatctgcaaccagtacgacgtcgccctgtccatcggcgacggcctgcgccccggctccatctacgacgccaacgacacggcccagttcgccgagctgctgacccagggcgagctgacgcgccgcgcgtgggagaaggacgtgcaggtgatgaacgagggccccggccacgtgcccatgcacaagatccccgagaacatgcagaagcagctggagtggtgcaacgaggcgcccttctacaccctgggccccctgacgaccgacatcgcgcccggctacgaccacatcacctccgccatcggcgcggccaacatcggcgccctgggcaccgccctgctgtgctacgtgacgcccaaggagcacctgggcctgcccaaccgcgacgacgtgaaggcgggcgtcatcgcctacaagatcgccgcccacgcggccgacctggccaagcagcacccccacgcccaggcgtgggacgacgcgctgtccaaggcgcgcttcgagttccgctggatggaccagttcgcgctgtccctggaccccatgacggcgatgtccttccacgacgagacgctgcccgcggacggcgcgaaggtcgcccacttctgctccatgtgcggccccaagttctgctccatgaagatcacggaggacatccgcaagtacgccgaggagaacggctacggctccgccgaggaggccatccgccagggcatggacgccatgtccgaggagttcaacatcgccaagaagacgatctccggcgagcagcacggcgaggtcggcggcgagatctacctgcccgagtcctacgtc

cgcacgcatccaacgaccgtatacgcatcgtccaatgaccgtcggtgtcctctctgcctccgttttgtgagatgtctcaggcttggtgcatcctcgggtggccagccacgttgcgcgtcgtgctgcttgcctctcttgcgcctctgtggtactggaaaatatcatcgaggcccgtttttttgctcccatttcctttccgctacatcttgaaagcaaacgacaaacgaagcagcaagcaaagagcacgaggacggtgaacaagtctgtcacctgtatacatctatttccccgcgggtgcacctactctctctcctgccccggcagagtcagctgccttacgtgacggatcccgcgtctcgaacagagcgcgcagaggaacgctgaaggtctcgcctctgtcgcacctcagcgcggcatacaccacaataaccacctgacgaatgcgcttggttcttcgtccattagcgaagcgtccggttcacacacgtgccacgttggcgaggtggcaggtgacaatgatcggtgga

gaggagcgcaaggccaccggctaccgcgagttctccggccgccacgagttcccctccaacaccatgcacgccctgctggccatgggcatctggctgggcgccatccacttcaacgccctgctgctgctgttctccttcctgttcctgcccttctccaagttcctggtggtgttcggcctgctgctgctgttcatgatcctgcccatcgacccctactccaagttcggccgccgcctgtcccgctacatctccaagcacgcctgctcctacttccccatcaccctgcacgtggaggacatccacgccttccaccccgaccgcgcctacgtgttcggcttcgagccccactccgtgctgcccatcggcgtggtggccctggccgacctgaccggcttcatgcccctgcccaagatcaaggtgctggcctcctccgccgtgttctacacccccttcctgcgccacatctggacctggctgggcctgacccccgccaccaagaagaacttctcctccctgctggacgccggctactcctgcatcctggtgcccggcggcgtgcaggagaccttccacatggagcccggctccgagatcgccttcctgcgcgcccgccgcggcttcgtgcgcatcgccatggagatgggctcccccctggtgcccgtgttctgcttcggccagtcccacgtgtacaagtggtggaagcccggcggcaagttctacctgcagttctcccgcgccatcaagttcacccccatcttcttctggggcatcttcggctcccccctgccctaccagcaccccatgcacgtggtggtgggcaagcccatcgacgtgaagaagaacccccagcccatcgtggaggaggtgatcgaggtgcacgaccgcttcgtggaggccctgcaggacctgttcgagcgccacaaggcccaggtgggc

gctggtcgtgtgatggactgttgccgccacacttgctgccttgacctgtgaatatccctgccgcttttatcaaacagcctcagtgtgtttgatcttgtgtgtacgcgcttttgcgagttgctagctgcttgtgctatttgcgaataccacccccagcatccccttccctcgtttcatatcgcttgcatcccaaccgcaacttatctacgctgtcctgctatccctcagcgctgctcctgctcctgctcactgcccctcgcacagccttggtttgggctccgcctgtattctcctggtactgcaacctgtaaaccagcactgcaatgctgatgcacgggaagtagtgggatgggaacacaaatggacttaaggatctaagtaagattcgaagcgctcgaccgtgccggacggactgcagccccatgtcgtagtgaccgccaatgtaagtgggctggcgtttccctgtacgtgagtcaacgtcactgcacgcgcaccaccctctcgaccggcaggaccaggcatcgcgagat

gcgacctgcgtcgctcggcgggctccgggccccggcgcccagcgaggcccctccccgtgcgcg ggcgcgccatccccccccgcatcatcgtggtgtcctcctcctcctccaaggtgaaccccctgaagaccgaggccgtggtgtcctccggcctggccgaccgcctgcgcctgggctccctgaccgaggacggcctgtcctacaaggagaagttcatcgtgcgctgctacgaggtgggcatcaacaagaccgccaccgtggagaccatcgccaacctgctgcaggaggtgggctgcaaccacgcccagtccgtgggctactccaccgccggcttctccaccacccccaccatgcgcaagctgcgcctgatctgggtgaccgcccgcatgcacatcgagatctacaagtaccccgcctggtccgacgtggtggagatcgagtcctggggccagggcgagggcaagatcggcacccgccgcgactggatcctgcgcgactacgccaccggccaggtgatcggccgcgccacctccaagtgggtgatgatgaaccaggacacccgccgcctgcagaaggtggacgtggacgtgcgcgacgagtacctggtgcactgcccccgcgagctgcgcctggccttccccgaggagaacaactcctccctgaagaagatctccaagctggaggacccctcccagtactccaagctgggcctggtgccccgccgcgccgacctggacatgaaccagcacgtgaacaacgtgacctacatcggctgggtgctggagtccatgccccaggagatcatcgacacccacgagctgcagaccatcaccctggactaccgccgcgagtgccagcacgacgacgtggtggactccctgacctcccccgagccctccgaggacgccgaggccgtgttcaaccacaacggcaccaacggctccgccaacgtgtccgccaacgaccacggctgccgcaacttcctgcacctgctgcgcctgtccggcaacggcctggagatcaaccgcggccgcaccgagtggcgcaagaagcccacccgcATGGACTACAA

ggagcgacgagtgtgcgtgcggggctggcgggagtgggacgccctcctcgctcctctctgttctgaacggaacaatcggccaccccgcgctacgcgccacgcatcgagcaacgaagaaaaccccccgatgataggttgcggtggctgccgggatatagatccggccgcacatcaaagggcccctccgccagagaagaagctcctttcccagcagactccttctgctgccaaaacacttctctgtccacagcaacaccaaaggatgaacagatcaacttgcgtctccgcgtagcttcctcggctagcgtgcttgcaacaggtccctgcactattatcttcctgctttcctctgaattatgcggcaggcgagcgctcgctctggcgagcgctccttcgcgccgccctcgctgatcgagtgtacagtcaatgaatggtgagctcctcactcagcgcgcctgcgcggggatgcggaacgccgccgccgccttgtcttttgcacgcgcgactccgtcgcttcgcgggtggcacccccattgaaaaaaacctcaattctgtttgtggaagacacggtgtacccccaaccacccacctgcacctctattattggtattattgacgcgggagcgggcgttgtactctacaacgtagcgtctctggttttcagctggctcccaccattgtaaattcttgctaaaatagtgcgtggttatgtgagaggtatggtgtaacagggcgtcagtcatgttggttttcgtgctgatctcgggcacaaggcgtcgtcgacgtgacgtgcccgtgatgagagcaataccgcgctcaaagccgacgcatggcctttactccgcactccaaacgactgtcgctcgtatttttcggatatctattttttaagagcgagcacagcgccgggcatgggcctgaaaggcctcgcggccgtgctcgtggtgggggccgcgagcgcgtggggcatcgcggcagtgcaccaggcgcagacggaggaacgcatggtgagtgcgcatcacaagatgcatgtcttgttgtctgtactataatgctagagcatcaccaggggcttagtcatcgcacctgctttggtcattacagaaattgcacaagggcgtcctccgggatgaggagatgtaccagctcaagctggagcggcttcgagccaagcaggagcgcggcgcatgacgacctacccacatgcgaa gagc Nucleotide sequence of transforming DNA contained in pSZ6377GarmFATA1(G108A) SEQ ID NO: 130 gctcttcgcgaaggtcattttccagaacaacgaccatggcttgtcttagcgatcgctcgaatgactgctagtgagtcgtacgctcgacccagtcgctcgcaggagaacgcggcaactgccgagcttcggcttgccagtcgtgactcgtatgtgatcaggaatcattggcattggtagcattataattcggcttccgcgctgtttatgggcatggcaatgtctcatgcagtcgaccttagtcaaccaattctgggtggccagctccgggcgaccgggctccgtgtcgccgggcaccacctcctgccatgagtaacagggccgccctctcctcccgacgttggccaactgaataccgtgtcttggggccctacatgatgggctgcctagtcgggcgggacgcgcaactgcccgcgcaatctgggacgtggtctgaatcctccaggcgggtttccccgagaaagaaagggtgccgatttcaaagcagagccatgtgccgggccctgtggcctgtgttggcgcctatgtagtcaccccccctcacccaattgtcgccagtttgcgcaatccataaactcaaaactgcagcttctgagctgcgctgttcaa

ctgatgtccgtggtctgcaacaacaagaaccactccgcccgccccaagctgcccaactcctccctgctgcccggcttcgacgtggtggtccaggccgcggccacccgcttcaagaaggagacgacgaccacccgcgccacgctgacgttcgacccccccacgaccaactccgagcgcgccaagcagcgcaagcacaccatcgacccctcctcccccgacttccagcccatcccctccttcgaggagtgcttccccaagtccacgaaggagcacaaggaggtggtgcacgaggagtccggccacgtcctgaaggtgcccttccgccgcgtgcacctgtccggcggcgagcccgccttcgacaactacgacacgtccggcccccagaacgtcaacgcccacatcggcctggcgaagctgcgcaaggagtggatcgaccgccgcgagaagctgggcacgccccgctacacgcagatgtactacgcgaagcagggcatcatcacggaggagatgctgtactgcgcgacgcgcgagaagctggaccccgagttcgtccgctccgaggtcgcgcggggccgcgccatcatcccctccaacaagaagcacctggagctggagcccatgatcgtgggccgcaagttcctggtgaaggtgaacgcgaacatcggcaactccgccgtggcctcctccatcgaggaggaggtctacaaggtgcagtgggccaccatgtggggcgccgacaccatcatggacctgtccacgggccgccacatccacgagacgcgcgagtggatcctgcgcaactccgcggtccccgtgggcaccgtccccatctaccaggcgctggagaaggtggacggcatcgcggagaacctgaactgggaggtgttccgcgagacgctgatcgagcaggccgagcagggcgtggactacttcacgatccacgcgggcgtgctgctgcgctacatccccctgaccgccaagcgcctgacgggcatcgtgtcccgcggcggctccatccacgcgaagtggtgcctggcctaccacaaggagaacttcgcctacgagcactgggacgacatcctggacatctgcaaccagtacgacgtcgccctgtccatcggcgacggcctgcgccccggctccatctacgacgccaacgacacggcccagttcgccgagctgctgacccagggcgagctgacgcgccgcgcgtgggagaaggacgtgcaggtgatgaacgagggccccggccacgtgcccatgcacaagatccccgagaacatgcagaagcagctggagtggtgcaacgaggcgcccttctacaccctgggccccctgacgaccgacatcgcgcccggctacgaccacatcacctccgccatcggcgcggccaacatcggcgccctgggcaccgccctgctgtgctacgtgacgcccaaggagcacctgggcctgcccaaccgcgacgacgtgaaggcgggcgtcatcgcctacaagatcgccgcccacgcggccgacctggccaagcagcacccccacgcccaggcgtgggacgacgcgctgtccaaggcgcgcttcgagttccgctggatggaccagttcgcgctgtccctggaccccatgacggcgatgtccttccacgacgagacgctgcccgcggacggcgcgaaggtcgcccacttctgctccatgtgcggccccaagttctgctccatgaagatcacggaggacatccgcaagtacgccgaggagaacggctacggctccgccgaggaggccatccgccagggcatggacgccatgtccgaggagttcaacatcgccaagaagacgatctccggcgagcagcacggcgaggtcggcggcgagatctacctgcccgagtcctacgtc

cgcacgcatccaacgaccgtatacgcatcgtccaatgaccgtcggtgtcctctctgcctccgttttgtgagatgtctcaggcttggtgcatcctcgggtggccagccacgttgcgcgtcgtgctgcttgcctctcttgcgcctctgtggtactggaaaatatcatcgaggcccgtttttttgctcccatttcctttccgctacatcttgaaagcaaacgacaaacgaagcagcaagcaaagagcacgaggacggtgaacaagtctgtcacctgtatacatctatttccccgcgggtgcacctactctctctcctgccccggcagagtcagctgccttacgtgacggatcccgcgtctcgaacagagcgcgcagaggaacgctgaaggtctcgcctctgtcgcacctcagcgcggcatacaccacaataaccacctgacgaatgcgcttggttcttcgtccattagcgaagcgtccggttcacacacgtgccacgttggcgaggtggcaggtgacaatgatcggtgga

ccgcatccactttctcggcgttcaatgcccgctgcggcgacctgcgtcgctcggcgggctccgggccccggcgcccagcgaggcccctccccgtgcgcg ggcgcgccatccccccccgcatcatcgtggtgtcctcctcctcctccaaggtgaaccccctgaagaccgaggccgtggtgtcctccggcctggccgaccgcctgcgcctgggctccctgaccgaggacggcctgtcctacaaggagaagttcatcgtgcgctgctacgaggtgggcatcaacaagaccgccaccgtggagaccatcgccaacctgctgcaggaggtgggctgcaaccacgcccagtccgtgggctactccaccgccggcttctccaccacccccaccatgcgcaagctgcgcctgatctgggtgaccgcccgcatgcacatcgagatctacaagtaccccgcctggtccgacgtggtggagatcgagtcctggggccagggcgagggcaagatcggcacccgccgcgactggatcctgcgcgactacgccaccggccaggtgatcggccgcgccacctccaagtgggtgatgatgaaccaggacacccgccgcctgcagaaggtggacgtggacgtgcgcgacgagtacctggtgcactgcccccgcgagctgcgcctggccttccccgaggagaacaactcctccctgaagaagatctccaagctggaggacccctcccagtactccaagctgggcctggtgccccgccgcgccgacctggacatgaaccagcacgtgaacaacgtgacctacatcggctgggtgctggagtccatgccccaggagatcatcgacacccacgagctgcagaccatcaccctggactaccgccgcgagtgccagcacgacgacgtggtggactccctgacctcccccgagccctccgaggacgccgaggccgtgacaaccacaacggcaccaacggctccgccaacgtgtccgccaacgaccacggctgccgcaacttcctgcacctgctgcgcctgtccggcaacggcctggagatcaaccgcggccgcaccgagtggcgcaagaagcccacccgcATGGACTACAAGGACCACGACGGCGACTACAAGGACCACGACATCGACTACA

ctctctgttctgaacggaacaatcggccaccccgcgctacgcgccacgcatcgagcaacgaagaaaaccccccgatgataggttgcggtggctgccgggatatagatccggccgcacatcaaagggcccctccgccagagaagaagctcctttcccagcagactccttctgctgccaaaacacttctctgtccacagcaacaccaaaggatgaacagatcaacttgcgtctccgcgtagcttcctcggctagcgtgcttgcaacaggtccctgcactattatcttcctgctttcctctgaattatgcggcaggcgagcgctcgctctggcgagcgctccttcgcgccgccctcgctgatcgagtgtacagtcaatgaatggtgagctcctcactcagcgcgcctgcgcggggatgcggaacgccgccgccgccttgtcttttgcacgcgcgactccgtcgcttcgcgggtggcacccccattgaaaaaaacctcaattctgtttgtggaagacacggtgtacccccaaccacccacctgcacctctattattggtattattgacgcgggagcgggcgttgtactctacaacgtagcgtctctggttttcagctggctcccaccattgtaaattcttgctaaaatagtgcgtggttatgtgagaggtatggtgtaacagggcgtcagtcatgttggttttcgtgctgatctcgggcacaaggcgtcgtcgacgtgacgtgcccgtgatgagagcaataccgcgctcaaagccgacgcatggcctttactccgcactccaaacgactgtcgctcgtatttttcggatatctattttttaagagcgagcacagcgccgggcatgggcctgaaaggcctcgcggccgtgctcgtggtgggggccgcgagcgcgtggggcatcgcggcagtgcaccaggcgcagacggaggaacgcatggtgagtgcgcatcacaagatgcatgtcttgttgtctgtactataatgctagagcatcaccaggggcttagtcatcgcacctgctttggtcattacagaaattgcacaagggcgtcctccgggatgaggagatgtaccagctcaagctggagcggcttcgagccaagcaggagcgcggcgcatgacgacctacccacatgc gaagagcNucleotide sequence of transforming DNA contained in pSZ6315 BnOTESEQ ID NO: 131 caccggcgcgctgcttcgcgtgccgggtgcagcaatcagatccaagtctgacgacttgcgcgcacgcgccggatccttcaattccaaagtgtcgtccgcgtgcgcttcttcgccttcgtcctcttgaacatccagcgacgcaagcgcagggcgctgggcggctggcgtcccgaaccggcctcggcgcacgcggctgaaattgccgatgtcggcaatgtagtgccgctccgcccacctctcaattaagtttttcagcgcgtggttgggaatgatctgcgctcatggggcgaaagaaggggttcagaggtgctttattgttactcgactgggcgtaccagcattcgtgcatgactgattatacatacaaaagtacagctcgcttcaatgccctgcgattcctactcccgagcgagcactcctctcaccgtcgggttgcttcccacgaccacgccggtaagagggtctgtggcctcgcgcccctcgcgagcgcatattccagccacgtctgtatgattttgcgctcatacgtctggcccgtcgaccccaaaatgacgggatcctgcataatatcgcccgaaatgggatccaggcattcgtcaggaggcgtcagccccgcgggagatgccggtcccgccgcattggaaaggtgtagagggggt

gcctgggcctgacgccccagatgggctgggacaactggaacacgttcgcctgcgacgtctccgagcagctgctgctggacacggccgaccgcatctccgacctgggcctgaaggacatgggctacaagtacatcatcctggacgactgctggtcctccggccgcgactccgacggcttcctggtcgccgacgagcagaagttccccaacggcatgggccacgtcgccgaccacctgcacaacaactccttcctgttcggcatgtactcctccgcgggcgagtacacgtgcgccggctaccccggctccctgggccgcgaggaggaggacgcccagttcttcgcgaacaaccgcgtggactacctgaagtacgacaactgctacaacaagggccagttcggcacgcccgagatctcctaccaccgctacaaggccatgtccgacgccctgaacaagacgggccgccccatcttctactccctgtgcaactggggccaggacctgaccttctactggggctccggcatcgcgaactcctggcgcatgtccggcgacgtcacggcggagttcacgcgccccgactcccgctgcccctgcgacggcgacgagtacgactgcaagtacgccggcttccactgctccatcatgaacatcctgaacaaggccgcccccatgggccagaacgcgggcgtcggcggctggaacgacctggacaacctggaggtcggcgtcggcaacctgacggacgacgaggagaaggcgcacttctccatgtgggccatggtgaagtcccccctgatcatcggcgcgaacgtgaacaacctgaaggcctcctcctactccatctactcccaggcgtccgtcatcgccatcaaccaggactccaacggcatccccgccacgcgcgtctggcgctactacgtgtccgacacggacgagtacggccagggcgagatccagatgtggtccggccccctggacaacggcgaccaggtcgtggcgctgctgaacggcggctccgtgtcccgccccatgaacacgaccctggaggagatcttcttcgactccaacctgggctccaagaagctgacctccacctgggacatctacgacctgtgggcgaaccgcgtcgacaactccacggcgtccgccatcctgggccgcaacaagaccgccaccggcatcctgtacaacgccaccgagcagtcctacaaggacggcctgtccaagaacgacacccgcctgttcggccagaagatcggctccctgtc

accggcgctgatgtggcgcggacgccgtcgtactctttcagactttactcttgaggaattgaacctttctcgcttgctggcatgtaaacattggcgcaattaattgtgtgatgaagaaagggtggcacaagatggatcgcgaatgtacgagatcgacaacgatggtgattgttatgaggggccaaacctggctcaatcttgtcgcatgtccggcgcaatgtgatccagcggcgtgactctcgcaacctggtagtgtgtgcgcaccgggtcgctttgattaaaactgatcgcattgccatcccgtcaactcacaagcctactctagctcccattgcgcactcgggcgcccggctcgatcaatgttctgagcggagggcgaagcgtcaggaaatcgtctcggcagctggaagcgcatggaatgcggagcggagatcgaatcaggatcccgcgtctcgaacagagcgcgcagaggaacgctgaaggtctcgcctctgtcgcacctcagcgcggcatacaccacaataaccacctgacgaatgcgcttggttcttcgtccattagcgaagcgt

tcgctcctctctgttctgaacggaacaatcggccaccccgcgctacgcgccacgcatcgagcaacgaagaaaaccccccgatgataggttgcggtggctgccgggatatagatccggccgcacatcaaagggcccctccgccagagaagaagctcctttcccagcagactccttctgctgccaaaacacttctctgtccacagcaacaccaaaggatgaacagatcaacttgcgtctccgcgtagcttcctcggctagcgtgcttgcaacaggtccctgcactattatcttcctgctttcctctgaattatgcggcaggcgagcgctcgctctggcgagcgctccttcgcgccgccctcgctgatcgagtgtacagtcaatgaatggtgagctccgcgcctgcgcgaggacgcagaacaacgctgccgccgtgtcttttgcacgcgcgactccggcgcttcgctggtggcacccccataaagaaaccctcaattctgtttgtggaagacacggtgtacccccacccacccacctgcacctctattattggtattattgacgcgggagtgggcgttgtaccctacaacgtagcttctctagttttcagctggctcccaccattgtaaattcatgctagaatagtgcgtggttatgtgagaggtatagtgtgtctgagcagacggggcgggatgcatgtcgtggtggtgatctttggctcaaggcgtcgtcgacgtgacgtgcccgatcatgagagcaataccgcgctcaaagccgacgcatagcctttactccgcaatccaaacgactgtcgctcgtatttfttggatatctattttaaagagcgagcacagcgccgggcatgggcctgaaaggcctcgcggccgtgctcgtggtgggggccgcgagcgcgtggggcatcgcggcagtgcaccaggcgcagacggaggaacgcatggtgcgtgcgcaatataagatacatgtattgttgt cctgcaggNucleotide sequence of BnOTE (D124A) in pSZ6316: SEQ ID NO: 132

Nucleotide sequence of BnOTE (D209A) in pSZ6317: SEQ ID NO: 133

Nucleotide sequence of BnOTE (D124A, D209A) in pSZ6318 SEQ ID NO: 134

Nucleotide sequence of transforming DNA contained in pSZ5083 GarmGATA1SEQ ID NO: 135ccctcaactgcgacgctgggaaccttctccgggcaggcgatgtgcgtgggtttgcctccttggcacggctctacaccgtcgagtacgccatgaggcggtgatggctgtgtcggttgccacttcgtccagagacggcaagtcgtccatcctctgcgtgtgtggcgcgacgctgcagcagtccctctgcagcagatgagcgtgactttggccatttcacgcactcgagtgtacacaatccatttttcttaaagcaaatgactgctgattgaccagatactgtaacgctgatttcgctccagatcgcacagatagcgaccatgttgctgcgtctgaaaatctggattccgaattcgaccctggcgctccatccatgcaacagatggcgacacttgttacaattcctgtcacccatcggcatggagcaggtccacttagattcccgatcacccacgcacatctcgctaatagtcattcgttcgtgtcttcgatcaatctcaagtgagtgtgcatggatcttggttgacgatgcggtatgggtttgcgccgctggctgcagggtctgcccaaggcaagctaacccagctcctctccccgacaatactctcgcaggcaaagccggtcacttgccttccagattgccaataaactcaattatggcctctgtcatgccatccatgggtctgatgaatggtcacgctcgtgtcctgaccgttccccagcctctggcgtcccctgccccgcccaccagcccacgccgcgcggcagtcgctgccaaggctgtctcggaGGTACC CTTTCTTGCGCTATGACACTTCCAGCAAAAGGTAGGGCGGGCTGCGAGACGGCTTCCCGGCGCTGCATGCAACACCGATGATGCTTCGACCCCCCGAAGCTCCTTCGGGGCTGCATGGGCGCTCCGATGCCGCTCCAGGGCGAGCGCTGTTTAAATAGCCAGGCCCCCGATTGCAAAGACATTATAGCGAGCTACCAAAGCCATATTCAAACACCTAGATCACTACCACTTCTACACAGGCCACTCGAGCTTGTGATCGCACTCCGCTAAGGGGGCGCCTCTTCCTCTTCGTTTCAGTCACAACCCGCAAAC TCTAGAATATC Aatgatcgagcaggacggcctccacgccggctcccccgccgcctgggtggagcgcctgttcggctacgactgggcccagcagaccatcggctgctccgacgccgccgtgttccgcctgtccgcccagggccgccccgtgctgttcgtgaagaccgacctgtccggcgccctgaacgagctgcaggacgaggccgcccgcctgtcctggctggccaccaccggcgtgccctgcgccgccgtgctggacgtggtgaccgaggccggccgcgactggctgctgctgggcgaggtgcccggccaggacctgctgtcctcccacctggcccccgccgagaaggtgtccatcatggccgacgccatgcgccgcctgcacaccctggaccccgccacctgccccttcgaccaccaggccaagcaccgcatcgagcgcgcccgcacccgcatggaggccggcctggtggaccaggacgacctggacgaggagcaccagggcctggcccccgccgagctgttcgcccgcctgaaggcccgcatgcccgacggcgaggacctggtggtgacccacggcgacgcctgcctgcccaacatcatggtggagaacggccgcttctccggcttcatcgactgcggccgcctgggcgtggccgaccgctaccaggacatcgccctggccacccgcgacatcgccgaggagctgggcggcgagtgggccgaccgcttcctggtgctgtacggcatcgccgcccccgactcccagcgcatcgccttctaccgcctgctggacgagttcttctga CAATTGACGCCCGCGCGGCGCACCTGACCTGTTCTCTCGAGGGCGCCTGTTCTGCCTTGCGAAACAAGCCCCTGGAGCATGCGTGCATGATCGTCTCTGGCGCCCCGCCGCGCGGTTTGTCGCCCTCGCGGGCGCCGCGGCCGCGGGGGCGCATTGAAATTGTTGCAAACCCCACCTGACAGATTGAGGGCCCAGGCAGGAAGGCGTTGAGATGGAGGTACAGGAGTCAAGTAACTGAAAGTTTTTATGATAACTAACAACAAAGGGTCGTTTCTGGCCAGCGAATGACAAGAACAAGATTCCACATTTCCGTGTAGAGGCTTGCCATCGAATGTGAGCGGGCGGGCCGCGGACCCGACAAAACCCTTACGACGTGGTAAGAAAAACGTGGCGGGCACTGTCCCTGTAGCCTGAAGACCAGCAGGAGACGATCGGAAGCATCACAGCACAGGATCCCGCGTCTCGAACAGAGCGCGCAGAGGAACGCTGAAGGTCTCGCCTCTGTCGCACCTCAGCGCGGCATACACCACAATAACCACCTGACGAATGCGCTTGGTTCTTCGTCCATTAGCGAAGCGTCCGGTTCACACACGTGCCACGTTGGCGAGGTGGCAGGTGACAATGATCGGTGGAGCTGATGGTCGAAACGTTCACAGCCTAGGGATATC GTGAAAACTCGCTCGACCGCCCGCGTCCCGCAGGCAGCGATGACGTGTGCGTGACCTGGGTGTTTCGTCGAAAGGCCAGCAACCCCAAATCGCAGGCGATCCGGAGATTGGGATCTGATCCGAGCTTGGACCAGATCCCCCACGATGCGGCACGGGAACTGCATCGACTCGGCGCGGAACCCAGCTTTCGTAAATGCCAGATTGGTGTCCGATACCTTGATTTGCCATCAGCGAAACAAGACTTCAGCAGCGAGCGTATTTGGCGGGCGTGCTACCAGGGTTGCATACATTGCCCATTTCTGTCTGGACCGCTTTACCGGCGCAGAGGGTGAGTTGATGGGGTTGGCAGGCATCGAAACGCGCGTGCATGGTGTGTGTGTCTGTTTTCGGCTGCACAATTTCAATAGTCGGATGGGCGACGGTAGAATTGGGTGTTGCGCTCGCGTGCATGCCTCGCCCCGTCGGGTGTCATGACCGGGACTGGAATCCCCCCTCGCGACCCTCCTGCTAACGCTCCCGACTCTCCCGCCCGCGCGCAGGATAGACTCTAGTTCAACCAATCGACA ACTAGT atggccaccgcatccactttctcggcgttcaatgcccgctgcggcgacctgcgtcgctcggcgggctccgggccccggcgcccagcgaggcccctccccgtgcgcgggcgcgccatccccccccgcatcatcgtggtgtcctcctcctcctccaaggtgaaccccctgaagaccgaggccgtggtgtcctccggcctggccgaccgcctgcgcctgggctccctgaccgaggacggcctgtcctacaaggagaagttcatcgtgcgctgctacgaggtgggcatcaacaagaccgccaccgtggagaccatcgccaacctgctgcaggaggtgggctgcaaccacgcccagtccgtgggctactccaccggcggcttctccaccacccccaccatgcgcaagctgcgcctgatctgggtgaccgcccgcatgcacatcgagatctacaagtaccccgcctggtccgacgtggtggagatcgagtcctggggccagggcgagggcaagatcggcacccgccgcgactggatcctgcgcgactacgccaccggccaggtgatcggccgcgccacctccaagtgggtgatgatgaaccaggacacccgccgcctgcagaaggtggacgtggacgtgcgcgacgagtacctggtgcactgcccccgcgagctgcgcctggccttccccgaggagaacaactcctccctgaagaagatctccaagctggaggacccctcccagtactccaagctgggcctggtgccccgccgcgccgacctggacatgaaccagcacgtgaacaacgtgacctacatcggctgggtgctggagtccatgccccaggagatcatcgacacccacgagctgcagaccatcaccctggactaccgccgcgagtgccagcacgacgacgtggtggactccctgacctcccccgagccctccgaggacgccgaggccgtgttcaaccacaacggcaccaacggctccgccaacgtgtccgccaacgaccacggctgccgcaacttcctgcacctgctgcgcctgtccggcaacggcctggagatcaaccgcggccgcaccgagtggcgcaagaagcccacccgcatggactacaaggaccacgacggcgactacaaggaccacgacatcgactacaaggacgacgacgacaagtga ATCGATgcagcagcagctcggatagtatcgacacactctggacgctggtcgtgtgatggactgttgccgccacacttgctgccttgacctgtgaatatccctgccgcttttatcaaacagcctcagtgtgtttgatcttgtgtgtacgcgcttttgcgagttgctagctgcttgtgctatttgcgaataccacccccagcatccccttccctcgtttcatatcgcttgcatcccaaccgcaacttatctacgctgtcctgctatccctcagcgctgctcctgctcctgctcactgcccctcgcacagccttggtttgggctccgcctgtattctcctggtactgcaacctgtaaaccagcactgcaatgctgatgcacgggaagtagtgggatgggaacacaaatggaAAGCTTGAGCTCcagcgccatgccacgccctttgatggcttcaagtacgattacggtgttggattgtgtgtttgttgcgtagtgtgcatggtttagaataatacacttgatttcttgctcacggcaatctcggcttgtccgcaggttcaaccccatttcggagtctcaggtcagccgcgcaatgaccagccgctacttcaaggacttgcacgacaacgccgaggtgagctatgtttaggacttgattggaaattgtcgtcgacgcatattcgcgctccgcgacagcacccaagcaaaatgtcaagtgcgttccgatttgcgtccgcaggtcgatgttgtgatcgtcggcgccggatccgccggtctgtcctgcgcttacgagctgaccaagcaccctgacgtccgggtacgcgagctgagattcgattagacataaattgaagattaaacccgtagaaaaatttgatggtcgcgaaactgtgctcgattgcaagaaattgatcgtcctccactccgcaggtcgccatcatcgagcagggcgttgctcccggcggcggcgcctggctggggggacagctgttctcggccatgtgtgtacgtagaaggatgaatttcagctggttttcgttgcacagctgtttgtgcatgatttgtttcagactattgttgaatgtttttagatttcttaggatgcatgatttgtctgcatgcgactAmino acid sequence of Gm FATA wild-type parental gene; D3997, pSZ5083.SEQ ID NO: 136MATASTFSAFNARCGDLRRSAGSGPRRPARPLPVRGRAIPPRIIVVSSSSSKVNPLKTEAVVSSGLADRLRLGSLTEDGLSYKEKFIVRCYEVGINKTATVETIANLLQEVGCNHAQSVGYSTGGFSTTPTMRKLRLIWVTARMHIEIYKYPAWSDVVEIESWGQGEGKIGTRRDWILRDYATGQVIGRATSKWVMMNQDTRRLQKVDVDVRDEYLVHCPRELRLAFPEENNSSLKKISKLEDPSQYSKLGLVPRRADLDMNQHVNNVTYIGWVLESMPQEIIDTHELQTITLDYRRECQHDDVVDSLTSPEPSEDAEAVFNHNGTNGSANVSANDHGCRNFLHLLRLSGNGLEINRGRTEWRKKPTRMDYKDHDGDYKDHDIDYKDDDDKAmino acid sequence of Gm FATA S111A, V193A mutant gene; D3998, pSZ5084.SEQ ID NO: 137MATASTFSAFNARCGDLRRSAGSGPRRPARPLPVRGRAIPPRIIVVSSSSSKVNPLKTEAVVSSGLADRLRLGSLTEDGLSYKEKFIVRCYEVGINKTATVETIANLLQEVGCNHAQSVGYSTGGFaTTPTMRKLRLIWVTARMHIEIYKYPAWSDVVEI ESWGQGEGKIGTRRDWILRDYATGQVIGRATSKWVMMNQDTRRLQKVDaDVRDEYLVHCPRELRLAFPEENNSSLKKISKLEDPSQYSKLGLVPRRADLDMNQHVNNVTYIGWVLESMPQEIIDTHELQTITLDYRRECQHDDVVDSLTSPEPSEDAEAVFNHNGTNGSANVSANDHGCRNFLHLLRLSGNGLEINRGRTEWRKKPTRMDYKDHDGDYKDHDIDYKDDDDKAmino acid sequence of Gm FATA S111V, V193A mutant gene; D3999, pSZ5085.SEQ ID NO: 138MATASTFSAFNARCGDLRRSAGSGPRRPARPLPVRGRAIPPRIIVVSSSSSKVNPLKTEAVVSSGLADRLRLGSLTEDGLSYKEKFIVRCYEVGINKTATVETIANLLQEVGCNHAQSVGYSTGGFvTTPTMRKLRLIWVTARMHIEIYKYPAWSDVVEI ESWGQGEGKIGTRRDWILRDYATGQVIGRATSKWVMMNQDTRRLQKVDaDVRDEYLVHCPRELRLAFPEENNSSLKKISKLEDPSQYSKLGLVPRRADLDMNQHVNNVTYIGWVLESMPQEIIDTHELQTITLDYRRECQHDDVVDSLTSPEPSEDAEAVFNHNGTNGSANVSANDHGCRNFLHLLRLSGNGLEINRGRTEWRKKPTRMDYKDHDGDYKDHDIDYKDDDDK Amino acid sequence of Gm FATA G96A mutant gene; D4000,pSZ5086. SEQ ID NO: 139MATASTFSAFNARCGDLRRSAGSGPRRPARPLPVRGRAIPPRIIVVSSSSSKVNPLKTEAVVSSGLADRLRLGSLTEDGLSYKEKFIVRCYEVGINKTATVETIANLLQEVaCNHAQSVGYSTGGFSTTPTMRKLRLIWVTARMHIEIYKYPAWSDVVEIESWGQGEGKIGTRRDWILRDYATGQVIGRATSKWVMMNQDTRRLQKVDVDVRDEYLVHCPRELRLAFPEENNSSLKKISKLEDPSQYSKLGLVPRRADLDMNQHVNNVTYIGWVLESMPQEIIDTHELQTITLDYRRECQHDDVVDSLTSPEPSEDAEAVFNHNGTNGSANVSANDHGCRNFLHLLRLSGNGLEINRGRTEWRKKPTRMDYKDHDGDYKDHDIDYKDDDDK Amino acid sequence of Gm FATA G96T mutant gene; D4001,pSZ5087. SEQ ID NO: 140MATASTFSAFNARCGDLRRSAGSGPRRPARPLPVRGRAIPPRIIVVSSSSSKVNPLKTEAVVSSGLADRLRLGSLTEDGLSYKEKFIVRCYEVGINKTATVETIANLLQEVtCNHAQSVGYSTGGFSTTPTMRKLRLIWVTARMHIEIYKYPAWSDVVEI ESWGQGEGKIGTRRDWILRDYATGQVIGRATSKWVMMNQDTRRLQKVDVDVRDEYLVHCPRELRLAFPEENNSSLKKISKLEDPSQYSKLGLVPRRADLDMNQHVNNVTYIGWVLESMPQEIIDTHELQTITLDYRRECQHDDVVDSLTSPEPSEDAEAVFNHNGTNGSANVSANDHGCRNFLHLLRLSGNGLEINRGRTEWRKKPTRMDYKDHDGDYKDHDIDYKDDDDK Amino acid sequence of Gm FATA G96V mutant gene; D4002,pSZ5088. SEQ ID NO: 141MATASTFSAFNARCGDLRRSAGSGPRRPARPLPVRGRAIPPRIIVVSSSSSKVNPLKTEAVVSSGLADRLRLGSLTEDGLSYKEKFIVRCYEVGINKTATVETIANLLQEVvCNHAQSVGYSTGGFSTTYPAWSDVVEIESWGQGEGKIGTRRDWILRDYATGQVIGRATSKWVMMNQDTRRLQKVDVDVRDEYLVHCPRELRLSKLEDPSQYSKLGLVPRRADLDMNQHVNNVTYIGWVLESMPQEIIDTHELQTITLDYRRECQHDDVVDSLTSPEPSEDAEAVFNHNGTNGSANVSANDHGCRNFLHLLRLSGNGLEINRGRTEWRKKPTRMDYKDHDGDYKDHDIDYKDDDDK Amino acid sequence of GmFATA G108A mutant gene; D4003,pSZ5089. SEQ ID NO: 142MATASTFSAFNARCGDLRRSAGSGPRRPARPLPVRGRAIPPRIIVVSSSSSKVNPLKTEAVVSSGLADRLRLGSLTEDGLSYKEKFIVRCYEVGINKTATVETIANLLQEVGCNHAQSVGYSTaGFSTTPTMRKLRLIWVTARMHIEIYKYPAWSDVVEIESWGQGEGKIGTRRDWILRDYATGQVIGRATSKWVMMNQDTRRLQKVDVDVRDEYLVHCPRELRLAFPEENNSSLKKISKLEDPSQYSKLGLVPRRADLDMNQHVNNVTYIGWVLESMPQEIIDTHELQTITLDYRRECQHDDVVDSLTSPEPSEDAEAVFNHNGTNGSANVSANDHGCRNFLHLLRLSGNGLEINRGRTEWRKKPTRMDYKDHDGDYKDHDIDYKDDDDK Amino acid sequence of GmFATA L91F mutant gene; D4004,pSZ5090. SEQ ID NO: 143MATASTFSAFNARCGDLRRSAGSGPRRPARPLPVRGRAIPPRIIVVSSSSSKVNPLKTEAVVSSGLADRLRLGSLTEDGLSYKEKFIVRCYEVGINKTATVETIANfLQEVGCDCNHAQSVGYSTGGFSTTPTMRKLRLIWVTARMHIEIYKYPAWSDVVEIESWGQGEGKIGTRRDWILRDYATGQVIGRATSKWVMMNQDTRRLQKVDVDVRDEYLVHCPRELRLAFPEENNSSLKKISKLEDPSQYSKLGLVPRRADLDMNQHVNNVTYIGWVLESMPQEIIDTHELQTITLDYRRECQHDDVVDSLTSPEPSEDAEAVFNHNGTNGSANVSANDHGCRNFLHLLRLSGNGLEINRGRTEWRKKPTRMDYKDHDGDYKDHDIDYKDDDDK Amino acid sequence of GmFATA L91K mutant gene; D4005,pSZ5091. SEQ ID NO: 144MATASTFSAFNARCGDLRRSAGSGPRRPARPLPVRGRAIPPRIIVVSSSSKVNPLKTEAVVSSGLADRLRLGSLTEDGLSYKEKFIVRCYEVGINKTATVETIANkLQEVGCNHAQSVGYSTGGFSTTPTMRKLRLIWVTARMHIEIYKYPAWSDVVEIESWGQGEGKIGTRRDWILRDYATGQVIGRATSKWVMMNQDTRRLQKVDVDVRDEYLVHCPRELRLAFPEENNSSLKKISKLEDPSQYSKLGLVPRRADLDMNQHVNNVTYIGWVLESMPQEIDTHELQTITLDYRRECQHDDVVDSLTSPEPSEDAEAVFNHNGTNGSANVSANDHGCRNFLHLLRLSGNGLEINRGRTEWRKKPTRMDYKDHDGDYKDHDIDYKDDDDKFigure 10. Amino acid sequence of GmFATA L91S mutant gene;D4006, pSZ5092. The algal transit peptide is underlined, the FLAG epitope tag isuppercase bold and the L91S residue is lower-case cold SEQ ID NO: 145MATASTFSAFNARCGDLRRSAGSGPRRPARPLPVRGRAIPPRIIVVSSSSSKVNPLKTEAVVSSGLADRLRLGSLTEDGLSYKEKFIVRCYEVGINKTATVETIANsLQEVGCNHAQSVGYSTGGFSTTPTMRKLRLIWVTARMHIEIYKYPAWSDVVEIESWGQGEGKIGTRRDWILRDYATGQVIGRATSKWVMMNQDTRRLQKVDVDVRDEYLVHCPRELRLAFPEENNSSLKKISKLEDPSQYSKLGLVPRRADLDMNQHVNNVTYIGWVLESMPQEIIDTHELQTITLDYRRECQHDDVVDSLTSPEPSEDAEAVFNHNGTNGSANVSANDHGCRNFLHLLRLSGNGLEINRGRTEWRKKPTRMDYKDHDGDYKDHDIDYKDDDDKAmino acid sequence of GmFATA G108V mutant gene; D4007, pSZ5093. The algaltransit peptide is underlined, the FLAG epitope tag is uppercase bold and the G108V residue islower-case bold. SEQ ID NO: 146MATASTFSAFNARCGDLRRSAGSGPRRPARPLPVRGRAIPPRIIVVSSSSSKVNPLKTEAVVSSGLADRLRLGSLTEDGLSYKEKFIVRCYEVGINKTATVETIANLLQEVGCNHAQSVGYSTvGFSTTPTMRKLRLIWVTARMHIEIYKYPAWSDVVEIESWGQGEGKIGTRRDWILRDYATGQVIGRATSKWVMMNQDTRRLQKVDVDVRDEYLVHCPRELRLAFPEENNSSLKKISKLEDPSQYSKLGLVPRRADLDMNQHVNNVTYIGWVLESMPQEIIDTHELQTITLDYRRECQHDDVVDSLTSPEPSEDAEAVFNHNGTNGSANVSANDHGCRNFLHLLRLSGNGLEINRGRTEWRKKPTRMDYKDHDGDYKDHDIDYKDDDDK Amino acid sequence of GmFATA T156F mutant gene; D4008,pSZ5094. The algal transit peptide is underlined, the FLAG epitope tag is uppercasebold and the T156F residue is lower-case bold. SEQ ID NO: 147MATASTFSAFNARCGDLRRSAGSGPRRPARPLPVRGRAIPPRIIVVSSSSSKVNPLKTEAVVSSGLADRLRLGSLTEDGLSYKEKFIVRCYEVGINKTATVETIANLLQEVGCNHAQSVGYSTGGFSTTPTMRKLRLIWVTARMHIEIYKYPAWSDVVEIESWGQGEGKIGfRRDWILRDYATGQVIGRATSKWVMMNQDTRRLQKVDVDVRDEYLVHCPRELRLAFPEENNSSLKKISKLEDPSQYSKLGLVPRRADLDMNQHVNNVTYIGWVLESMPQEIIDTHELQTITLDYRRECQHDDVVDSLTSPEPSEDAEAVFNHNGTNGSANVSANDHGCRNFLHLLRLSGNGLEINRGRTEWRKKPTRMDYKDHDGDYKDHDIDYKDDDDK Amino acid sequence of GmFATA T156A mutant gene; D4009,pSZ5095. The algal transit peptide is underlined, the FLAG epitope tag is uppercasebold and the T156A residue is lower-case bold. SEQ ID NO: 148MATASTFSAFNARCGDLRRSAGSGPRRPARPLPVRGRAIPPRIIVVSSSSSKVNPLKTEAVVSSGLADRLRLGSLTEDGLSYKEKFIVRCYEVGINKTATVETIANLLQEVGCNHAQSVGYSTGGFSTTPTMRKLRLIWVTARMHIEIYKYPAWSDVVEIESWGQGEGKIGaRRDWILRDYATGQVIGRATSKWVMMNQDTRRLQKVDVDVRDEYLVHCPRELRLAFPEENNSSLKKISKLEDPSQYSKLGLVPRRADLDMNQHVNNVTYIGWVLESMPQEIIDTHELQTITLDYRRECQHDDVVDSLTSPEPSEDAEAVFNHNGTNGSANVSANDHGCRNFLHLLRLSGNGLEINRGRTEWRKKPTRMDYKDHDGDYKDHDIDYKDDDDKAmino acid sequence of GmFATA T156K mutant gene; D4010, pSZ5096. Thealgal transit peptide is underlined, the FLAG epitope tag is uppercase bold and the T156K residueis lower-case bold. SEQ ID NO:  149MATASTFSAFNARCGDLRRSAGSGPRRPARPLPVRGRAIPPRIIVVSSSSSKVNPLKTEAVVSSGLADRLRLGSLTEDGLSYKEKFIVRCYEVGINKTATVETIANLLQEVGCNHAQSVGYSTGGFSTTPTMRKLRLIWVTARMHIEIYKYPAWSDVVEIESWGQGEGKIGkRRDWILRDYATGQVIGRATSKWVMMNQDTRRLQKVDVDVRDEYLVHCPRELRLAFPEENNSSLKKISKLEDPSQYSKLGLVPRRADLDMNQHVNNVTYIGWVLESMPQEIIDTHELQTITLDYRRECQHDDVVDSLTSPEPSEDAEAVFNHNGTNGSANVSANDHGCRNFLHLLRLSGNGLEINRGRTEWRKKPTRMDYKDHDGDYKDHDIDYKDDDDKAmino acid sequence of GmFATA T156V mutant gene; D4011, pSZ5097. Thealgal transit peptide is underlined, the FLAG epitope tag is uppercase bold and the T156V residueis lower-case bold. SEQ ID NO:  150MATASTFSAFNARCGDLRRSAGSGPRRPARPLPVRGRAIPPRIIVVSSSSSKVNPLKTEAVVSSGLADRLRLGSLTEDGLSYKEKFIVRCYEVGINKTATVETIANLLQEVGCNHAQSVGYSTGGFSTTPTMRKLRLIWVTARMHIEIYKYPAWSDVVEIESWGQGEGKIGvRRDWILRDYATGQVIGRATSKWVMMNQDTRRLQKVDVDVRDEYLVHCPRELRLAFPEENNSSLKKISKLEDPSQYSKLGLVPRRADLDMNQHVNNVTYIGWVLESMPQEIIDTHELQTITLDYRRECQHDDVVDSLTSPEPSEDAEAVFNHNGTNGSANVSANDHGCRNFLHLLRLSGNGLEINRGRTEWRKKPTRMDYKDHDGDYKDHDIDYKDDDDKNucleotide sequence of the GmFATA S111A, V193A mutant gene (D3998,pSZ5084). The promoter, 3′UTR, selection marker and targeting arms are the same as pSZ5083.SEQ ID NO: 151atggccaccgcatccactttctcggcgttcaatgcccgctgcggcgacctgcgtcgctcggcgggctccgggccccggcgcccagcgaggcccctccccgtgcgcgggcgcgccatccccccccgcatcatcgtggtgtcctcctcctcctccaaggtgaaccccctgaagaccgaggccgtggtgtcctccggcctggccgaccgcctgcgcctgggctccctgaccgaggacggcctgtcctacaaggagaagttcatcgtgcgctgctacgaggtgggcatcaacaagaccgccaccgtggagaccatcgccaacctgctgcaggaggtgggctgcaaccacgcccagtccgtgggctactccaccggcggcttcgccaccacccccaccatgcgcaagctgcgcctgatctgggtgaccgcccgcatgcacatcgagatctacaagtaccccgcctggtccgacgtggtggagatcgagtcctggggccagggcgagggcaagatcggcacccgccgcgactggatcctgcgcgactacgccaccggccaggtgatcggccgcgccacctccaagtgggtgatgatgaaccaggacacccgccgcctgcagaaggtggacgcggacgtgcgcgacgagtacctggtgcactgcccccgcgagctgcgcctggccttccccgaggagaacaactcctccctgaagaagatctccaagctggaggacccctcccagtactccaagctgggcctggtgccccgccgcgccgacctggacatgaaccagcacgtgaacaacgtgacctacatcggctgggtgctggagtccatgccccaggagatcatcgacacccacgagctgcagaccatcaccctggactaccgccgcgagtgccagcacgacgacgtggtggactccctgacctcccccgagccctccgaggacgccgaggccgtgttcaaccacaacggcaccaacggctccgccaacgtgtccgccaacgaccacggctgccgcaacttcctgcacctgctgcgcctgtccggcaacggcctggagatcaaccgcggccgcaccgagtggcgcaagaagcccacccgcatggactacaaggaccacgacggcgactacaaggaccacgacatcgactacaaggacgacgacgacaagtgaNucleotide sequence of the GmFATA s111V, V193A mutant gene (D3999,pSZ5085). The promoter, 3′UTR, selection marker and targeting arms are the same as pSZ5083.SEQ ID NO: 152atggccaccgcatccactttctcggcgttcaatgcccgctgcggcgacctgcgtcgctcggcgggctccgggccccggcgcccagcgaggcccctccccgtgcgcgggcgcgccatccccccccgcatcatcgtggtgtcctcctcctcctccaaggtgaaccccctgaagaccgaggccgtggtgtcctccggcctggccgaccgcctgcgcctgggctccctgaccgaggacggcctgtcctacaaggagaagttcatcgtgcgctgctacgaggtgggcatcaacaagaccgccaccgtggagaccatcgccaacctgctgcaggaggtgggctgcaaccacgcccagtccgtgggctactccaccggcggcttcgtcaccacccccaccatgcgcaagctgcgcctgatctgggtgaccgcccgcatgcacatcgagatctacaagtaccccgcctggtccgacgtggtggagatcgagtcctggggccagggcgagggcaagatcggcacccgccgcgactggatcctgcgcgactacgccaccggccaggtgatcggccgcgccacctccaagtgggtgatgatgaaccaggacacccgccgcctgcagaaggtggacgcggacgtgcgcgacgagtacctggtgcactgcccccgcgagctgcgcctggccttccccgaggagaacaactcctccctgaagaagatctccaagctggaggacccctcccagtactccaagctgggcctggtgccccgccgcgccgacctggacatgaaccagcacgtgaacaacgtgacctacatcggctgggtgctggagtccatgccccaggagatcatcgacacccacgagctgcagaccatcaccctggactaccgccgcgagtgccagcacgacgacgtggtggactccctgacctcccccgagccctccgaggacgccgaggccgtgttcaaccacaacggcaccaacggctccgccaacgtgtccgccaacgaccacggctgccgcaacttcctgcacctgctgcgcctgtccggcaacggcctggagatcaaccgcggccgcaccgagtggcgcaagaagcccacccgcatggactacaaggaccacgacggcgactacaaggaccacgacatcgactacaaggacgacgacgacaagtgaNucleotide sequence of the GmFATA G96A mutant gene (D4000, pSZ5086). Thepromoter, 3′UTR, selection marker and targeting arms are the same as pSZ5083SEQ ID NO: 153atggccaccgcatccactttctcggcgttcaatgcccgctgcggcgacctgcgtcgctcggcgggctccgggccccggcgcccagcgaggcccctccccgtgcgcgggcgcgccatccccccccgcatcatcgtggtgtcctcctcctcctccaaggtgaaccccctgaagaccgaggccgtggtgtcctccggcctggccgaccgcctgcgcctgggctccctgaccgaggacggcctgtcctacaaggagaagttcatcgtgcgctgctacgaggtgggcatcaacaagaccgccaccgtggagaccatcgccaacctgctgcaggaggtggcgtgcaaccacgcccagtccgtgggctactccaccggcggcttctccaccacccccaccatgcgcaagctgcgcctgatctgggtgaccgcccgcatgcacatcgagatctacaagtaccccgcctggtccgacgtggtggagatcgagtcctggggccagggcgagggcaagatcggcacccgccgcgactggatcctgcgcgactacgccaccggccaggtgatcggccgcgccacctccaagtgggtgatgatgaaccaggacacccgccgcctgcagaaggtggacgtggacgtgcgcgacgagtacctggtgcactgcccccgcgagctgcgcctggccttccccgaggagaacaactcctccctgaagaagatctccaagctggaggacccctcccagtactccaagctgggcctggtgccccgccgcgccgacctggacatgaaccagcacgtgaacaacgtgacctacatcggctgggtgctggagtccatgccccaggagatcatcgacacccacgagctgcagaccatcaccctggactaccgccgcgagtgccagcacgacgacgtggtggactccctgacctcccccgagccctccgaggacgccgaggccgtgttcaaccacaacggcaccaacggctccgccaacgtgtccgccaacgaccacggctgccgcaacttcctgcacctgctgcgcctgtccggcaacggcctggagatcaaccgcggccgcaccgagtggcgcaagaagcccacccgcatggactacaaggaccacgacggcgactacaaggaccacgacatcgactacaaggacgacgacgacaagtgaNucleotide sequence of the GmFATA G96T mutant gene (D4001, pSZ5087). Thepromoter, 3′UTR, selection marker and targeting arms are the same as pSZ5083SEQ ID NO: 154atggccaccgcatccactttctcggcgttcaatgcccgctgcggcgacctgcgtcgctcggcgggctccgggccccggcgcccagcgaggcccctccccgtgcgcgggcgcgccatccccccccgcatcatcgtggtgtcctcctcctcctccaaggtgaaccccctgaagaccgaggccgtggtgtcctccggcctggccgaccgcctgcgcctgggctccctgaccgaggacggcctgtcctacaaggagaagttcatcgtgcgctgctacgaggtgggcatcaacaagaccgccaccgtggagaccatcgccaacctgctgcaggaggtgacgtgcaaccacgcccagtccgtgggctactccaccggcggcttctccaccacccccaccatgcgcaagctgcgcctgatctgggtgaccgcccgcatgcacatcgagatctacaagtaccccgcctggtccgacgtggtggagatcgagtcctggggccagggcgagggcaagatcggcacccgccgcgactggatcctgcgcgactacgccaccggccaggtgatcggccgcgccacctccaagtgggtgatgatgaaccaggacacccgccgcctgcagaaggtggacgtggacgtgcgcgacgagtacctggtgcactgcccccgcgagctgcgcctggccttccccgaggagaacaactcctccctgaagaagatctccaagctggaggacccctcccagtactccaagctgggcctggtgccccgccgcgccgacctggacatgaaccagcacgtgaacaacgtgacctacatcggctgggtgctggagtccatgccccaggagatcatcgacacccacgagctgcagaccatcaccctggactaccgccgcgagtgccagcacgacgacgtggtggactccctgacctcccccgagccctccgaggacgccgaggccgtgttcaaccacaacggcaccaacggctccgccaacgtgtccgccaacgaccacggctgccgcaacttcctgcacctgctgcgcctgtccggcaacggcctggagatcaaccgcggccgcaccgagtggcgcaagaagcccacccgcatggactacaaggaccacgacggcgactacaaggaccacgacatcgactacaaggacgacgacgacaagtgaNucleotide sequence of the GmFATA G96V mutant gene (D4002, pSZ5088). Thepromoter, 3′UTR, selection marker and targeting arms are the same as pSZ5083SEQ ID NO: 155atggccaccgcatccactttctcggcgttcaatgcccgctgcggcgacctgcgtcgctcggcgggctccgggccccggcgcccagcgaggcccctccccgtgcgcgggcgcgccatccccccccgcatcatcgtggtgtcctcctcctcctccaaggtgaaccccctgaagaccgaggccgtggtgtcctccggcctggccgaccgcctgcgcctgggctccctgaccgaggacggcctgtcctacaaggagaagttcatcgtgcgctgctacgaggtgggcatcaacaagaccgccaccgtggagaccatcgccaacctgctgcaggaggtggtgtgcaaccacgcccagtccgtgggctactccaccggcggcttctccaccacccccaccatgcgcaagctgcgcctgatctgggtgaccgcccgcatgcacatcgagatctacaagtaccccgcctggtccgacgtggtggagatcgagtcctggggccagggcgagggcaagatcggcacccgccgcgactggatcctgcgcgactacgccaccggccaggtgatcggccgcgccacctccaagtgggtgatgatgaaccaggacacccgccgcctgcagaaggtggacgtggacgtgcgcgacgagtacctggtgcactgcccccgcgagctgcgcctggccttccccgaggagaacaactcctccctgaagaagatctccaagctggaggacccctcccagtactccaagctgggcctggtgccccgccgcgccgacctggacatgaaccagcacgtgaacaacgtgacctacatcggctgggtgctggagtccatgccccaggagatcatcgacacccacgagctgcagaccatcaccctggactaccgccgcgagtgccagcacgacgacgtggtggactccctgacctcccccgagccctccgaggacgccgaggccgtgttcaaccacaacggcaccaacggctccgccaacgtgtccgccaacgaccacggctgccgcaacttcctgcacctgctgcgcctgtccggcaacggcctggagatcaaccgcggccgcaccgagtggcgcaagaagcccacccgcatggactacaaggaccacgacggcgactacaaggaccacgacatcgactacaaggacgacgacgacaagtgaNucleotide sequence of the GmFATA G108A mutant gene (D4003, pSZ5089). Thepromoter, 3′UTR, selection marker and targeting arms are the same as pSZ50836.SEQ ID NO: 156atggccaccgcatccactttctcggcgttcaatgcccgctgcggcgacctgcgtcgctcggcgggctccgggccccggcgcccagcgaggcccctccccgtgcgcgggcgcgccatccccccccgcatcatcgtggtgtcctcctcctcctccaaggtgaaccccctgaagaccgaggccgtggtgtcctccggcctggccgaccgcctgcgcctgggctccctgaccgaggacggcctgtcctacaaggagaagttcatcgtgcgctgctacgaggtgggcatcaacaagaccgccaccgtggagaccatcgccaacctgctgcaggaggtgggctgcaaccacgcccagtccgtgggctactccaccgccggcttctccaccacccccaccatgcgcaagctgcgcctgatctgggtgaccgcccgcatgcacatcgagatctacaagtaccccgcctggtccgacgtggtggagatcgagtcctggggccagggcgagggcaagatcggcacccgccgcgactggatcctgcgcgactacgccaccggccaggtgatcggccgcgccacctccaagtgggtgatgatgaaccaggacacccgccgcctgcagaaggtggacgtggacgtgcgcgacgagtacctggtgcactgcccccgcgagctgcgcctggccttccccgaggagaacaactcctccctgaagaagatctccaagctggaggacccctcccagtactccaagctgggcctggtgccccgccgcgccgacctggacatgaaccagcacgtgaacaacgtgacctacatcggctgggtgctggagtccatgccccaggagatcatcgacacccacgagctgcagaccatcaccctggactaccgccgcgagtgccagcacgacgacgtggtggactccctgacctcccccgagccctccgaggacgccgaggccgtgttcaaccacaacggcaccaacggctccgccaacgtgtccgccaacgaccacggctgccgcaacttcctgcacctgctgcgcctgtccggcaacggcctggagatcaaccgcggccgcaccgagtggcgcaagaagcccacccgcatggactacaaggaccacgacggcgactacaaggaccacgacatcgactacaaggacgacgacgacaagtgaNucleotide sequence of the GmFATA L91F mutant gene (D4004, pSZ5090). Thepromoter, 3′UTR, selection marker and targeting arms are the same as pSZ5083SEQ ID NO: 157atggccaccgcatccactttctcggcgttcaatgcccgctgcggcgacctgcgtcgctcggcgggctccgggccccggcgcccagcgaggcccctccccgtgcgcgggcgcgccatccccccccgcatcatcgtggtgtcctcctcctcctccaaggtgaaccccctgaagaccgaggccgtggtgtcctccggcctggccgaccgcctgcgcctgggctccctgaccgaggacggcctgtcctacaaggagaagttcatcgtgcgctgctacgaggtgggcatcaacaagaccgccaccgtggagaccatcgccaacttcctgcaggaggtgggctgcaaccacgcccagtccgtgggctactccaccggcggcttctccaccacccccaccatgcgcaagctgcgcctgatctgggtgaccgcccgcatgcacatcgagatctacaagtaccccgcctggtccgacgtggtggagatcgagtcctggggccagggcgagggcaagatcggcacccgccgcgactggatcctgcgcgactacgccaccggccaggtgatcggccgcgccacctccaagtgggtgatgatgaaccaggacacccgccgcctgcagaaggtggacgtggacgtgcgcgacgagtacctggtgcactgcccccgcgagctgcgcctggccttccccgaggagaacaactcctccctgaagaagatctccaagctggaggacccctcccagtactccaagctgggcctggtgccccgccgcgccgacctggacatgaaccagcacgtgaacaacgtgacctacatcggctgggtgctggagtccatgccccaggagatcatcgacacccacgagctgcagaccatcaccctggactaccgccgcgagtgccagcacgacgacgtggtggactccctgacctcccccgagccctccgaggacgccgaggccgtgttcaaccacaacggcaccaacggctccgccaacgtgtccgccaacgaccacggctgccgcaacttcctgcacctgctgcgcctgtccggcaacggcctggagatcaaccgcggccgcaccgagtggcgcaagaagcccacccgcatggactacaaggaccacgacggcgactacaaggaccacgacatcgactacaaggacgacgacgacaagtgaNucleotide sequence of the GmFATA L91K mutant gene (D4005, pSZ5091).SEQ ID NO: 158atggccaccgcatccactttctcggcgttcaatgcccgctgcggcgacctgcgtcgctcggcgggctccgggccccggcgcccagcgaggcccctccccgtgcgcgggcgcgccatccccccccgcatcatcgtggtgtcctcctcctcctccaaggtgaaccccctgaagaccgaggccgtggtgtcctccggcctggccgaccgcctgcgcctgggctccctgaccgaggacggcctgtcctacaaggagaagttcatcgtgcgctgctacgaggtgggcatcaacaagaccgccaccgtggagaccatcgccaacaagctgcaggaggtgggctgcaaccacgcccagtccgtgggctactccaccggcggcttctccaccacccccaccatgcgcaagctgcgcctgatctgggtgaccgcccgcatgcacatcgagatctacaagtaccccgcctggtccgacgtggtggagatcgagtcctggggccagggcgagggcaagatcggcacccgccgcgactggatcctgcgcgactacgccaccggccaggtgatcggccgcgccacctccaagtgggtgatgatgaaccaggacacccgccgcctgcagaaggtggacgtggacgtgcgcgacgagtacctggtgcactgcccccgcgagctgcgcctggccttccccgaggagaacaactcctccctgaagaagatctccaagctggaggacccctcccagtactccaagctgggcctggtgccccgccgcgccgacctggacatgaaccagcacgtgaacaacgtgacctacatcggctgggtgctggagtccatgccccaggagatcatcgacacccacgagctgcagaccatcaccctggactaccgccgcgagtgccagcacgacgacgtggtggactccctgacctcccccgagccctccgaggacgccgaggccgtgttcaaccacaacggcaccaacggctccgccaacgtgtccgccaacgaccacggctgccgcaacttcctgcacctgctgcgcctgtccggcaacggcctggagatcaaccgcggccgcaccgagtggcgcaagaagcccacccgcatggactacaaggaccacgacggcgactacaaggaccacgacatcgactacaaggacgacgacgacaagtgaNucleotide sequence of the GmFATA L91S mutant gene (D4006, pSZ5092).SEQ ID NO: 159atggccaccgcatccactttctcggcgttcaatgcccgctgcggcgacctgcgtcgctcggcgggctccgggccccggcgcccagcgaggcccctccccgtgcgcgggcgcgccatccccccccgcatcatcgtggtgtcctcctcctcctccaaggtgaaccccctgaagaccgaggccgtggtgtcctccggcctggccgaccgcctgcgcctgggctccctgaccgaggacggcctgtcctacaaggagaagttcatcgtgcgctgctacgaggtgggcatcaacaagaccgccaccgtggagaccatcgccaactcgctgcaggaggtgggctgcaaccacgcccagtccgtgggctactccaccggcggcttctccaccacccccaccatgcgcaagctgcgcctgatctgggtgaccgcccgcatgcacatcgagatctacaagtaccccgcctggtccgacgtggtggagatcgagtcctggggccagggcgagggcaagatcggcacccgccgcgactggatcctgcgcgactacgccaccggccaggtgatcggccgcgccacctccaagtgggtgatgatgaaccaggacacccgccgcctgcagaaggtggacgtggacgtgcgcgacgagtacctggtgcactgcccccgcgagctgcgcctggccttccccgaggagaacaactcctccctgaagaagatctccaagctggaggacccctcccagtactccaagctgggcctggtgccccgccgcgccgacctggacatgaaccagcacgtgaacaacgtgacctacatcggctgggtgctggagtccatgccccaggagatcatcgacacccacgagctgcagaccatcaccctggactaccgccgcgagtgccagcacgacgacgtggtggactccctgacctcccccgagccctccgaggacgccgaggccgtgttcaaccacaacggcaccaacggctccgccaacgtgtccgccaacgaccacggctgccgcaacttcctgcacctgctgcgcctgtccggcaacggcctggagatcaaccgcggccgcaccgagtggcgcaagaagcccacccgcatggactacaaggaccacgacggcgactacaaggaccacgacatcgactacaaggacgacgacgacaagtgaNucleotide sequence of the GmFATA G108V mutant gene (D4007, pSZ5093).SEQ ID NO: 160atggccaccgcatccactttctcggcgttcaatgcccgctgcggcgacctgcgtcgctcggcgggctccgggccccggcgcccagcgaggcccctccccgtgcgcgggcgcgccatccccccccgcatcatcgtggtgtcctcctcctcctccaaggtgaaccccctgaagaccgaggccgtggtgtcctccggcctggccgaccgcctgcgcctgggctccctgaccgaggacggcctgtcctacaaggagaagttcatcgtgcgctgctacgaggtgggcatcaacaagaccgccaccgtggagaccatcgccaacctgctgcaggaggtgggctgcaaccacgcccagtccgtgggctactccaccgtcggcttctccaccacccccaccatgcgcaagctgcgcctgatctgggtgaccgcccgcatgcacatcgagatctacaagtaccccgcctggtccgacgtggtggagatcgagtcctggggccagggcgagggcaagatcggcacccgccgcgactggatcctgcgcgactacgccaccggccaggtgatcggccgcgccacctccaagtgggtgatgatgaaccaggacacccgccgcctgcagaaggtggacgtggacgtgcgcgacgagtacctggtgcactgcccccgcgagctgcgcctggccttccccgaggagaacaactcctccctgaagaagatctccaagctggaggacccctcccagtactccaagctgggcctggtgccccgccgcgccgacctggacatgaaccagcacgtgaacaacgtgacctacatcggctgggtgctggagtccatgccccaggagatcatcgacacccacgagctgcagaccatcaccctggactaccgccgcgagtgccagcacgacgacgtggtggactccctgacctcccccgagccctccgaggacgccgaggccgtgttcaaccacaacggcaccaacggctccgccaacgtgtccgccaacgaccacggctgccgcaacttcctgcacctgctgcgcctgtccggcaacggcctggagatcaaccgcggccgcaccgagtggcgcaagaagcccacccgcatggactacaaggaccacgacggcgactacaaggaccacgacatcgactacaaggacgacgacgacaagtgaNucleotide sequence of the GmFATA T156F mutant gene (D4008, pSZ5094).SEQ ID NO: 161atggccaccgcatccactttctcggcgttcaatgcccgctgcggcgacctgcgtcgctcggcgggctccgggccccggcgcccagcgaggcccctccccgtgcgcgggcgcgccatccccccccgcatcatcgtggtgtcctcctcctcctccaaggtgaaccccctgaagaccgaggccgtggtgtcctccggcctggccgaccgcctgcgcctgggctccctgaccgaggacggcctgtcctacaaggagaagttcatcgtgcgctgctacgaggtgggcatcaacaagaccgccaccgtggagaccatcgccaacctgctgcaggaggtgggctgcaaccacgcccagtccgtgggctactccaccggcggcttctccaccacccccaccatgcgcaagctgcgcctgatctgggtgaccgcccgcatgcacatcgagatctacaagtaccccgcctggtccgacgtggtggagatcgagtcctggggccagggcgagggcaagatcggcttccgccgcgactggatcctgcgcgactacgccaccggccaggtgatcggccgcgccacctccaagtgggtgatgatgaaccaggacacccgccgcctgcagaaggtggacgtggacgtgcgcgacgagtacctggtgcactgcccccgcgagctgcgcctggccttccccgaggagaacaactcctccctgaagaagatctccaagctggaggacccctcccagtactccaagctgggcctggtgccccgccgcgccgacctggacatgaaccagcacgtgaacaacgtgacctacatcggctgggtgctggagtccatgccccaggagatcatcgacacccacgagctgcagaccatcaccctggactaccgccgcgagtgccagcacgacgacgtggtggactccctgacctcccccgagccctccgaggacgccgaggccgtgttcaaccacaacggcaccaacggctccgccaacgtgtccgccaacgaccacggctgccgcaacttcctgcacctgctgcgcctgtccggcaacggcctggagatcaaccgcggccgcaccgagtggcgcaagaagcccacccgcatggactacaaggaccacgacggcgactacaaggaccacgacatcgactacaaggacgacgacgacaagtgaNucleotide sequence of the GmFATA T156A mutant gene (D4009, pSZ5095)SEQ ID NO: 162atggccaccgcatccactttctcggcgttcaatgcccgctgcggcgacctgcgtcgctcggcgggctccgggccccggcgcccagcgaggcccctccccgtgcgcgggcgcgccatccccccccgcatcatcgtggtgtcctcctcctcctccaaggtgaaccccctgaagaccgaggccgtggtgtcctccggcctggccgaccgcctgcgcctgggctccctgaccgaggacggcctgtcctacaaggagaagttcatcgtgcgctgctacgaggtgggcatcaacaagaccgccaccgtggagaccatcgccaacctgctgcaggaggtgggctgcaaccacgcccagtccgtgggctactccaccggcggcttctccaccacccccaccatgcgcaagctgcgcctgatctgggtgaccgcccgcatgcacatcgagatctacaagtaccccgcctggtccgacgtggtggagatcgagtcctggggccagggcgagggcaagatcggcgcgcgccgcgactggatcctgcgcgactacgccaccggccaggtgatcggccgcgccacctccaagtgggtgatgatgaaccaggacacccgccgcctgcagaaggtggacgtggacgtgcgcgacgagtacctggtgcactgcccccgcgagctgcgcctggccttccccgaggagaacaactcctccctgaagaagatctccaagctggaggacccctcccagtactccaagctgggcctggtgccccgccgcgccgacctggacatgaaccagcacgtgaacaacgtgacctacatcggctgggtgctggagtccatgccccaggagatcatcgacacccacgagctgcagaccatcaccctggactaccgccgcgagtgccagcacgacgacgtggtggactccctgacctcccccgagccctccgaggacgccgaggccgtgttcaaccacaacggcaccaacggctccgccaacgtgtccgccaacgaccacggctgccgcaacttcctgcacctgctgcgcctgtccggcaacggcctggagatcaaccgcggccgcaccgagtggcgcaagaagcccacccgcatggactacaaggaccacgacggcgactacaaggaccacgacatcgactacaaggacgacgacgacaagtgaNucleotide sequence of the GmFATA T156K mutant gene (D4010, pSZ5096).SEQ ID NO: 163atggccaccgcatccactttctcggcgttcaatgcccgctgcggcgacctgcgtcgctcggcgggctccgggccccggcgcccagcgaggcccctccccgtgcgcgggcgcgccatccccccccgcatcatcgtggtgtcctcctcctcctccaaggtgaaccccctgaagaccgaggccgtggtgtcctccggcctggccgaccgcctgcgcctgggctccctgaccgaggacggcctgtcctacaaggagaagttcatcgtgcgctgctacgaggtgggcatcaacaagaccgccaccgtggagaccatcgccaacctgctgcaggaggtgggctgcaaccacgcccagtccgtgggctactccaccggcggcttctccaccacccccaccatgcgcaagctgcgcctgatctgggtgaccgcccgcatgcacatcgagatctacaagtaccccgcctggtccgacgtggtggagatcgagtcctggggccagggcgagggcaagatcggcaagcgccgcgactggatcctgcgcgactacgccaccggccaggtgatcggccgcgccacctccaagtgggtgatgatgaaccaggacacccgccgcctgcagaaggtggacgtggacgtgcgcgacgagtacctggtgcactgcccccgcgagctgcgcctggccttccccgaggagaacaactcctccctgaagaagatctccaagctggaggacccctcccagtactccaagctgggcctggtgccccgccgcgccgacctggacatgaaccagcacgtgaacaacgtgacctacatcggctgggtgctggagtccatgccccaggagatcatcgacacccacgagctgcagaccatcaccctggactaccgccgcgagtgccagcacgacgacgtggtggactccctgacctcccccgagccctccgaggacgccgaggccgtgttcaaccacaacggcaccaacggctccgccaacgtgtccgccaacgaccacggctgccgcaacttcctgcacctgctgcgcctgtccggcaacggcctggagatcaaccgcggccgcaccgagtggcgcaagaagcccacccgcatggactacaaggaccacgacggcgactacaaggaccacgacatcgactacaaggacgacgacgacaagtgaNucleotide sequence of the GmFATA T156V mutant gene (D4011, pSZ5097).SEQ ID NO: 164atggccaccgcatccactttctcggcgttcaatgcccgctgcggcgacctgcgtcgctcggcgggctccgggccccggcgcccagcgaggcccctccccgtgcgcgggcgcgccatccccccccgcatcatcgtggtgtcctcctcctcctccaaggtgaaccccctgaagaccgaggccgtggtgtcctccggcctggccgaccgcctgcgcctgggctccctgaccgaggacggcctgtcctacaaggagaagttcatcgtgcgctgctacgaggtgggcatcaacaagaccgccaccgtggagaccatcgccaacctgctgcaggaggtgggctgcaaccacgcccagtccgtgggctactccaccggcggcttctccaccacccccaccatgcgcaagctgcgcctgatctgggtgaccgcccgcatgcacatcgagatctacaagtaccccgcctggtccgacgtggtggagatcgagtcctggggccagggcgagggcaagatcggcgtgcgccgcgactggatcctgcgcgactacgccaccggccaggtgatcggccgcgccacctccaagtgggtgatgatgaaccaggacacccgccgcctgcagaaggtggacgtggacgtgcgcgacgagtacctggtgcactgcccccgcgagctgcgcctggccttccccgaggagaacaactcctccctgaagaagatctccaagctggaggacccctcccagtactccaagctgggcctggtgccccgccgcgccgacctggacatgaaccagcacgtgaacaacgtgacctacatcggctgggtgctggagtccatgccccaggagatcatcgacacccacgagctgcagaccatcaccctggactaccgccgcgagtgccagcacgacgacgtggtggactccctgacctcccccgagccctccgaggacgccgaggccgtgttcaaccacaacggcaccaacggctccgccaacgtgtccgccaacgaccacggctgccgcaacttcctgcacctgctgcgcctgtccggcaacggcctggagatcaaccgcggccgcaccgagtggcgcaagaagcccacccgcatggactacaaggaccacgacggcgactacaaggaccacgacatcgactacaaggacgacgacgacaagtgaAmino acid sequence of wild type BnOTE in pSZ6315 (See SEQ ID NO: 131)SEQ ID NO:  165MATASTFSAFNARCGDLRRSAGSGPRRPARPLPVRGRASQLRKPALDPLRAVISADQGSISPVNSCTPADRLRAGRLMEDGYSYKEKFIVRSYEVGINKTATVETIANLLQEVACNHVQKCGFSTDGFATTLTMRKLHLIWVTARMHIEIYKYPAWSDVVEIETWCQSEGRIGTRRDWILRDSATNEVIGRATSKWVMMNQDTRRLQRVTDEVRDEYLVFCPREPRLAFPEENNSSLKKIPKLEDPAQYSMLELKPRRADLDMNQHVNNVTYIGWVLESIPQEIIDTHELQVITLDYRRECQQDDIVDSLTTSEIPDDPISKFTGTNGSAMSSIQGHNESQFLHMLRLSENGQEINRGRTQWRKKSSRMDYKDHDGDYKDHDIDYKDDDDKAmino Acid sequence of BnOTE (D124A) in pSZ6316 (See SEQ ID NO: 132)SEQ ID NO: 166MATASTFSAFNARCGDLRRSAGSGPRRPARPLPVRGRASQLRKPALDPLRAVISADQGSISPVNSCTPADRLRAGRLMEDGYSYKEKFIVRSYEVGINKTATVETIANLLQEVACNHVQKCGFSTAGFATTLTMRKLHLIWVTARMHIEIYKYPAWSDVVEIETWCQSEGRIGTRRDWILRDSATNEVIGRATSKWVMMNQDTRRLQRVTDEVRDEYLVFCPREPRLAFPEENNSSLKKIPKLEDPAQYSMLELKPRRADLDMNQHVNNVTYIGWVLESIPQEIIDTHELQVITLDYRRECQQDDIVDSLTTSEIPDDPISKFTGTNGSAMSSIQGHNESQFLHMLRLSENGQEINRGRTQWRKKSSRMDYKDHDGDYKDHDIDYKDDDDKAmino Acid sequence of BnOTE (D209A) in pSZ6317 (See SEQ ID NO: 133)SEQ ID NO: 167MATASTFSAFNARCGDLRRSAGSGPRRPARPLPVRGRASQLRKPALDPLRAVISADQGSISPVNSCTPADRLRAGRLMEDGYSYKEKFIVRSYEVGINKTATVETIANLLQEVACNHVQKCGFSTDGFATTLTMRKLHLIWVTARMHIEIYKYPAWSDVVEIETWCQSEGRIGTRRDWILRDSATNEVIGRATSKWVMMNQDTRRLQRVTAEVRDEYLVFCPREPRLAFPEENNSSLKKIPKLEDPAQYSMLELKPRRADLDMNQHVNNVTYIGWVLESIPQEIIDTHELQVITLDYRRECQQDDIVDSLTTSEIPDDPISKFTGTNGSAMSSIQGHNESQFLHMLRLSENGQEINRGRTQWRKKSSRMDYKDHDGDYKDHDIDYKDDDDKAmino acid sequence of BnOTE (D124A, D209A) in pSZ6318 (See SEQ ID NO: 134)SEQ ID NO: 168MATASTFSAFNARCGDLRRSAGSGPRRPARPLPVRGRASQLRKPALDPLRAVISADQGSISPVNSCTPADRLRAGRLMEDGYSYKEKFIVRSYEVGINKTATVETIANLLQEVACNHVQKCGFSTAGFATTLTMRKLHLIWVTARMHIEIYKYPAWSDVVEIETWCQSEGRIGTRRDWILRDSATNEVIGRATSKWVMMNQDTRRLQRVTAEVRDEYLVFCPREPRLAFPEENNSSLKKIPKLEDPAQYSMLELKPRRADLDMNQHVNNVTYIGWVLESIPQEIIDTHELQVITLDYRRECQQDDIVDSLTTSEIPDDPISKFTGTNGSAMSSIQGHNESQFLHMLRLSENGQEINRGRTQWRKKSSRMDYKDHDGDYKDHDIDYKDDDDK CpauLPAAT SEQ ID NO: 169MAIPAAAVIFLFGLLFFTSGLIINLFQALCFVLVWPLSKNAYRRINRVFAELLLSELLCLFDWWAGAKLKLFTDPETFRLMGKEHALVIINHMTELDWMLGWVMGQHLGCLGS IL SVAKKS TKFLPVLGWSMWFSEYLYIERSWAKDRTTLKSHIERLTDYPLPFWMVIFVEGTRFTRTKLLAAQQYAASSGLPVPRNVLIPRTKGFVSCVSHMRSFVPAVYDVTVAFPKTSPPPTLLNLFEGQSIVLHVHIKRHAMKDLPESDDAVAQWCRDKFVEKDALLDKHNAEDTFSGQEVHRTGSRPIKSLLVVISWVVVITFGALKFLQWSSWKGKAFSVIGLGIVTLLMHMLILSSQAERSSNPAKVAQAKLKTELSISKKATDKEN CprocLPAAT1iSEQ ID NO: 170  MAIPAAAVIFLFGLIFFASGLIINLFQALCFVLIWPISKNAYRRINRVFAELLLSELLCLFDWWAGAKLKLFTDPETFRLMGKEHALVIINHMTELDWMVGWVMGQHFGCLGSILSVAKKSTKFLPVLGWSMWFTEYLYIERSWNKDKSTLKSHIERLKDYPLPFWLVIFAEGTRFTQTKLLAAQQYAASSGLPVPRNVLIPRTKGFVSCVSHMRSFVPAVYDLTVAFPKTSPPPTLLNLFEGQSVVLHVHIKRHAMKDLPESDDEVAQWCRDKFVEKDALLDKHNAEDTFSGQELQHTGRRPIKSLLVVISWVVVIAFGALKFLQWSSWKGKAFSVIGLGIVTLLMHMLILSSQAERSKPAKVAQAKLKTELSISKTVTDKEN CpcuLPAAT1SEQ ID NO: 171  MAIPSAAVVFLFGLLFFTSGLIINLFQAFCFVLISPLSKNAYRRINRVFAELLPLEFLWLFHWCAGAKLKLFTDPETFRLMGKEHALVIINHKIELDWMVGWVLGQHLGCLGS ISVAKKSTKFLPVFGWSLWFSGYLFLERSWAKDKITLKSHIESLKDYPLPFWLIIFVEGTRFTRTKLLAAQQYAASSGLPVPRNVLIPHTKGFVSSVSHMRSFVPAIYDVTVAFPKTSPPPTMLKLFEGQSVELHVHIKRHAMKDLPESDDAVAQWCRDKFVEKDALLDKHNSEDTFSGQEVHHVGRPIKALLVVISWVVVIIFGALKFLLWSSLLSSWKGKAFSVIGLGIVAGIVTLLMHILILSSQAEGSNPVKAAPAKLKTELSSSKK VTNKENChookLPAAT1 SEQ ID NO: 172MAIPSAAVVFLFGLLFFTSGLINLFQAFCFVLISPLSKNAYRRINRVFAELLPLEFLWLFHWCAGAKLKLFTDPETFRLMGKEHALVIINHKIELDWMVGWVLGQHLGCLGSILSVAKKSTKFLPVFGWSLWFSEYLFLERSWAKDKITLKSHIESLKDYPLPFWLIIFVEGTRFTRTKLLAAQQYAASSGLPVPRNVLIPHTKGFVSSVSHMRSFVPAIYDVTVAFPKTSPPPTMLKLFEGQSVELHVHIKRHAMKDLPESDDAVAQWCRDKFVEKDALLDKHNSEDTFSGQEVHHVGRPIKALLVVISWVVVIIFGALKFLLWSSLLSSWKGKAFSVIGLGIVAGIVTLLMHILILSSQAEGSNPVKAAPAKLKTELSSSKK VTNKENCignLPAAT1 SEQ ID NO: 173 MAIAAAAVI FLFGLLFFASGIIINLFQALCFVLIWPLSKNVYRRINRVFAELLLMDLLCLFHWWAGAKIKLFTDPETFRLMGMEHALVIMNHKIDLDWMVGWILGQHLGCLGSILSIAKKSTKFIPVLGWSVWFSEYLFLERSWAKDKSTLKSHMEKLKDYPLPFWLVIFVEGTRFTRTKLLAAQQYAASSGLPVPRNVLIPHTKGFVSCVSNMRSFVPAVYDVTVAFPKSSPPPTMLKLFEGQSIVLHVHIKRHALKDLPESDDAVAQWCRDKFVEKDALLDKHNAEDTFSGQEVHHIGRPIKSLLVVIAWVVVIIFGALKFLQWSSLLSTWKGKAFSVIGLGIATLLMHMLILSSQAERSNPAKVAK CavigLPAAT1 SEQ ID NO: 174MTIASAAVVFLFGILLFTSGLIINLFQAFCSVLVWPLSKNAYRRINRVFAEFLPLEFLWLFHWWAGAKLKLFTDPETFRLMGKEHALVIINHKIELDWMVGWVLGQHLGCLGSILSVAKKSTKFLPVFGWSLWFSEYLFLERNWAKDKKTLKSHIERLKDYPLPFWLIIFVEGTRFTRTKLLAAQQYAASAGLPVPRNVLIPHTKGFVSSVSHMRSFVPAIYDVTVAFPKTSPPPTMLKLFEGHFVELHVHIKRHAMKDLPESEDAVAQWCRDKFVEKDALLDKHNAEDTFSGQEVHHVGRPIKSLLVVISWVVVIIFGALKFLQWSSLLSSWKGIAFSVIGLGTVALLMQILILSSQAERSIPAKETPANLKTELSSSKKVTNK EN CavigLPAAT2SEQ ID NO: 175 MAIAAAAVIVPVSLLFFVSGLIVNLVQAVCFVLIRPLFKNTYRRINRVVAELWLELVWLIDWWAGVKIKVFTDHETFHLMGKEHALVICNHKSDIDWLVGWVLAQRSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYLFLERNWAKDESTLKSGLNRLKDYPLPFWLALFVEGTRFTRAKLLAAQQYAASSGLPVPRNVLIPRTKGFVSSVSHMRSFVPAIYDVTVAI PKTSPPPTLLRMFKGQSSVLHVHLKRHQMNDLPESDDAVAQWCRDIFVEKDALLDKHNAEDTFSGQELQDTGRPIKSLLIVISWAVLVVFGAVKFLQWSSLLSSWKGLAFSGIGLGVITLLMHILILFSQSERSTPAKVAPAKPKIEGESSKTEMEK EH CpalLPAAT1SEQ ID NO: 176  MAIAAAAVIVPLGLLFFVSGLIVNLVQAVCFVLIRPLSKNTYRRINRVVAELLWLELVWLIDWWAGVKIKVFTDHETLSLMGKEHALVICNHKSDIDWLVGWVLAQRSGCLGSTLAVMKKSSKFLPVIGWSMWFSEYLPESDDAVAQWCRDIFVEKDALLDKHNAEDTFSGQELQDTGRPIKSLLVVISWAVLVIFGAVKFLQWSSLLSSWKGLAFSGVGLGIITLLMHILILFSQSERSTPAKVAPAKPKKDGESSKTEIEKENVPGALLGQGREHPEVRPEPPEGLPPALLAGPVRGGHPLHPRQAAGRPAVRHLLRPARAPQRADPPHQGLRVLRVPHALLRARHLRRDRGHPQDLPPPHHAAHVQGPVLRAARAPEAPPDE GPCuPSR23 LPAAT2 SEQ ID NO: 177 MAIAAAAVI FLFGLIFFASGLIINLFQALCFVLIRPLSKNAYRRINRVFAELLLSELLCLFDWWAGAKLKLFTDPETFRLMGKEHALVIINHMTELDWMVGWVMGQHFGCLGSIISVAKKSTKFLPVLGWSMWFSEYLYLERSWAKDKSTLKSHIERLIDYPLPFWLVIFVEGTRFTRTKLLAAQQYAVSSGLPVPRNVLIPRTKGFVSCVSHMRSFVPAVYDVTVAFPKTSPPPTLLNLFEGQSIMLHVHIKRHAMKDLPESDDAVAEWCRDKFVEKDALLDKHNAEDTFSGQEVCHSGSRQLKSLLVVISWVVVTTFGALKFLQWSSWKGKAFSAIGLGIVTLLMHVLILSSQAERSNPAEVAQAKLKTGLSISKKVTDKEN CkoeLPAAT1SEQ ID NO: 178 MAI PAAVAVI PIGLLFIISGLIVNLIQAVVYVLIRPLSKNLHRKINKPIAELLWLELIWLVDWWAGIKVEVYADSQTLELMGKEHALLICNHRSDIDWLVGWVLAQRARCLGSALAIMKKSAKFLPVIGWSMWFSDYIFLDRTWAKDEKTLKSGFERLADFPMPFWLALFVEGTRFTKAKLLAAQEYAASRGLPVPQNVLIPRTKGFVTAVTHMRSYVPAIYDCTVDISKAHPAPSILRLIRGQSSVVKVQITRHSMQELPETADGISQWCMDLFVTKDGFLEKYHSKDIFGSLPVQNIGRPVKSLIVVLCWYCLMAFGLFKFFMWSSLLSSWEGILSLGLILLAVAIVMQILIQSTESERSTPVKSIQKDPSKETLLQN CkoeLPAAT2SEQ ID NO: 179 MHVLLEMVTFRFSSFFVFDNVQALCFVLIWPLSKSAYRKINRVFAELLLSELLCLFDWWAGAKLKLFTDPETFRLMGKEHALVITNHKIDLDWMIGWILGQHFGCLGSVISIAKKSTKFLPIFGWSLWFSEYLFLERNWAKDKRTLKSHIERMKDYPLPLWLILFVEGTRFTRTKLLAAQQYAASSGLPVPRNVLIPHTKGFVSSVSHMRSFVPAVYDVTVAFPKTSPPPTMLSLFEGQSVVLHVHIKRHAMKDLPDSDDAVAQWCRDKFVEKDALLDKHNAEDTFSGQEVHHVGRTIKSLLVVISWMVVIIFGALKFLQWSSLLSSWKGKAFSAIGLGIATLLMHVLVVFSQADRSNPAKVPPAKLNTELSSSKKVTNKEN CprocLPAAT2SEQ ID NO: 180 MAI PAAVAVIPIGLLFIISGLIVNLIQAVVYVLIRPLSKNLYRKINKPIAELLWLELIWLVDWWAGIKVEVYADSETLESMGKEHALLICNHRSDIDWLVGWVLAQRARCLGSALAIMKKSAKFLPVIGWSMWFSDYIFLDRTWEKDEKTLKSGFERLADFPMPFWLALFVEGTRFTKAKLLAAQEFAASRGLPVPQNVLIPRTKGFVTAVTHMRSYVPAIYDCTVDISKAHPAPSILRLIRGQSSVVKVQITRHSMQELPETPDGISQWCMDLFVTKDAFLEKYHSKDIFGSLPVHDIGRPVKSLIVVLCWYSLMAFGFYKFFMWSSLLSSWEGILSLGLVLIVIAIVMQILIQSSESERSTPVKSVQKDPSKETLLQN CavigGPAT9SEQ ID NO: 181 MATGGSLKPSSSDLDLDHPNIEDYLPSGSSINEPAGKLRLRDLLDISPTLTEAAGAIVDDSFTRCFKSIPREPWNWNLYLFPLWCIGVLIRYFILFPGRVIVLTMGWITVISSFIAVRVLLKGHDALQIKLERLIVQLLCSSFVASWTGVVKYHGPRPSIRPKQVYVANHT SMIDFFILDQMTVFSVIMQKHPGWVGLLQSTLLESVGCIWFDRAEAKDRGIVAKKLWDHVHGEGNNPLLIFPEGTCVNNNYSVMFKKGAFELGCTVCPVAIKYNKIFVDAFWNSKKQSFTRHLLQLMTSWAVVCDVWYLEPQTLKPGETPIEFAERVRDIISARAGLKKVPWDGYLKYSRPSPKHRERKQQTFAESVLQRLEE ChookGPAT9-1 SEQ ID NO: 182MATAGSLKPSRSELDFDRPNIEDYLPSGSSIIEPAGKLRLRDLLDISPTLTEAAGAIVDDSFTRCFKSNPPEPWNWNIYLFPLWCFGVLIRYLILFPARVIVLTIGWIIFLSSFIPVHLLLKGHDALRI KLERLLVELICSFFVASWTGVVKYHGPRPSIRPKQVYVANHTSMIDFFILDQMTVFSVIMQKHPGWVGLLQSTLLESVGCIWFDRAEAKDRGIVAKKLWDHVHGEGNNPLLIFPEGTCVNNNYSVMFKKGAFELGCTVCPVAIKYNKIFVDAFWNSKKQSFTRHLLQLMTSWAVVCDVWYLEPQTLKPGETPIEFAERVRDIISVRAGLKKVPWDGYLKYSRPSPKHTERKQQNFAESVLQRLEKK CignGPAT9-1 SEQ ID NO: 183MATGGRLKPSSSELDLDRANTEDYLPSGSSINEPVGKLRLRDLLDISPTLTEAAGAIVDDSFTRCFKSIPPEPWNWNIYLFPLWCFGVLIRYFILFPARVIVLTIGWITVISSFTAVRFLLKGHNALQIKLERLIVQLLCSSFVASWTGVVKYHGPRPSIRPKQVYVANHTSMIDFLILDQMTVFSVIMQKHPGWVGLLQSTLLESVGCIWFNRAEAKDREIVAKKLWDHVHGEGNNPLLIFPEGTCVNNHYSVMFKKGAFELGCTVCPVAIKYNKIFVDAFWNSRKQSFTMHLLQLMTSWAVVCDVWYLEPQTLKPGETAIEFAERVRDIIVRAGLKKVPWDGYLKYSRPSPKHRESKQQSFAESVLRRLEEK CignGPAT9-2 SEQ ID NO: 184MATGGRLKPSSSELDLDRANTEDYLPSGSSINEPVGKLRLRDLLDISPTLTEAAGAIVDDSFTRCFKSIPPEPWNWNIYLFPLWCFGVLIRYFILFPARVIVLTIGWITVISSFTAVRFLLKGHNALQIKLERLIVQLLCSSFVASWTGVVKYHGPRPSIRPKQVYVANHTSMIDFLILDQMTVFSVIMQKHPGWVGLLQSTLLESVGCIWFNRAEAKDREIVAKKLWDHVHGEGNNPLLI FPEGTCVNNHYSVMFKKGAFELGCTVCPVAIKYNKIFVDAFWNSKKHSFTRHLLQLMTSWAVVCDVWYLEPQTLKPGETPIEFAERVRDIISVRADLKKVPWDGYLKYSRPSPKHRERKQQKFAESVLRRLEEK CpalGPAT9-1 SEQ ID NO: 185MATAGRLKPSSSELELDLDRPNIEDYLPSGSSINEPAGKLRLRDLLDISPMLTEAAGAIVDDSFTRCFKSIPPEPWNWNIYLFPLWCFGVLIRYLILFPARVIVLTVGWITVISSFITVRFLLKGHDSLRIKLERLIVQLFCSSFVASWTGVVKYHGPRPSIRPQQVYVANHTSMIDFIILNQMTVFSAIMQKHPGWVGLIQSTILESVGCIWFNRAEAKDREIVAKKLLDHVHGEGNNPLLIFPEGTCVNNHYSVMFKKGAFELGCTVCPVAIKYNKIFVDAFWNSKKQSFTMHLLQLMTSWAVVCDVWYLEPQTLKPGETPIEFAERVRDIISVRAGLKKVPWDGYLKYSRPSPKHRERKQQSFAESVLRRLEKR CpalGPATt9-2SEQ ID NO: 186 MATAGRLKPSSSELELDLDRPNIEDYLPSGSSINEPAGKLRLRDLLDISPMLTEAAGAIVDDSFTRCFKSIPPEPWNWNIYLFPLWCFGVLIRYLILFPARVIVLTVGWITVISSFITVRFLLKGHDSLRIKLERLIVQLFCSSFVASWTGVVKYHGPRPSIRPQQVYVANHTSMIDFIILNQMTVFSAIMQKHPGWVGLIQSTILESVGCIWFNRAEAKDREIVAKKLLDHVHGEGNNPLLIFPEGTCVNNHYSVMFKKGAFELGCTVCPVAIKYNKIFVDAFWNSKKLSFTMHLLQLMTSWAVVCDVWYLEPQTLKPGETPIEFAERVRDIISVRAGLKKVPWDGYLKYSRPSPKHRERKQQTFAESVLRRLEEKGNVVPTVN CavigDGAT1SEQ ID NO: 187 MAIADGGIIGAAGSISALTADTDPPSLRRRNVPAGQASAVSAFSTESMAKHLCDPSREPSPSPKSSDDGKDPDIGSVDSLNEKPSSPAAGKGRLQHDLRFTYRASSPAHRKVKESPLSSSNIFKQSHAGLFNLCVVVLVAVNSRLIIENLMKYGLLIKTGFWFSSRSLRDWPLFMCCLSLPIFPLAAFLVEKLAQKNRLQEPTVVCCHVLITSVSILYPVLVILRCDSAVLSGVALMLFACIVWLKLVSYAHSNYDMRYVAKSLDKGEPVVDSVIADHPYRVDYKDLVYFMVAPTLCYQLSYPLTPCVRKSWIARQVMKLVLFTGVMGFIVEQYINP IVQNSKHPLKGDLLYAIERVLKLSVPNLYVWLCMFYCFFHLWLNILAELICFGDREFYKDWWNAKTVEEYWRMWNMPVHKWMVRHIYFPCLRNGIPRGVAVLIAFLVSAVFHELCIAVPCHVFKLWAFIGIMFQVPLVLVSNCLQKKFQSSMAGNMFFWFIFCIFGQPMCVLLYYHDLMNRKGSRID ChookDGAT1-1 SEQ ID NO: 188MAIADGGSAGAAGSISGSDPSPSTAPSLRRRNASAGQAFSTESMARDLCDPSREPSLSPKSSDDGKDPADDIGAADSVDSGGVKDEKPSSQAAAKARLEHDLRFTYRASSPAHRKVKESPLSSSNIFKQSHAGLFNLCVVVLVAVNSRLIIENLMKYGLLIKTGFWFSSRSLRDWPLFMCCLSLPIFPLAAFLVEKLAQKNRLQEPTVVCCHVIITSVSILYPVLVILRCDSAVLSGVALMLFACIVWLKLVSYAHANYDMRSVAKSLDKGETVADSVIVDHPYRVDYKDLVYFMVAPTLCYQLSYPLTPYVRKSWVARQVMKLVLFTGVMGFIVEQYINPIVQNSKHPLKGDLLYAIERVLKLSVPNLYVWLCMFYCFFHLWLNILAELTCFGDREFYKDWWNAKTVEEYWRMWNMPVHKWMVRHIYFPCLRNGIPRGVAVLIAFLVSAVFHELCIAVPCHVFKLWAFIGIMFQVPLVLVSNCLQKKFQSSMAGNMFFWFIFCIFGQPMCVLLYYHDLMNRKGSRID CavigLPCAT SEQ ID NO: 189MGLVSVAAAIGVSVPVARFLLCFLATIPVSFLWRLVPGRLPKHLYSAASGAILSYLSFGASSNLHFIVPMTLGYLSMLFFRPFSGLLTFFLGFGYLIGCHVYYMSGDAWKEGGIDATGALMVLTLKVISCSMNYNDGLLKEEGLRESQKKNRLTKMPSLIEYFGYCLCCGSHFAGPVYEMKDYLEWTEGKGIWSRSQKEPKPSPFGGALRAIIQAAVCMAMYLYLVPHHPLTRFTEPVYYEWGFFRRLSYQYMAALTARWKYYFIWSISEASLIISGLGFSGWTESSPPKPRWDRAKNVDIIGVEFAKSSVQLPLVWNIQVSIWLRHYVYDRLVQNGKRPGFFQLLATQTVSAVWHGLYPGYIIFFVQSALMIAGSRVIYRWQQAVPPKMGLVKNIFVFFNFAYTLLVLNYSAVGFMVLSMHETLASYGSVYYIGTILPITLILLSYVIKPGKPARSKAHKEQ CpalLPCAT SEQ ID NO: 190MELGSVAAAIGVSVPVARFLLCFLATIPVSFLWRLVPGRLPKHLYSAASGAILSYLSFGPSSNLHFIVPMTLGYLSMLFFRFSGLLTFFLGFGYLIGCHVYYMSGDAWKEGGIDATGALMVLTLKVISCSINYNDGLLKEEGLRESQKKNRLTKMPSLIEYIGYCLCCGSHFAGPVYEMKDYLEWTEGKGVWSHSEKEPKPSPFGGALRAIIQAAVCMAMYMYLVPHHPLSRFTEPVYYEWGFFRRLSYQYMAGLTARWKYYFIWSISEASLIISGLGFSGWTESSPPKPRWDRAKNVDIIGVEFAKSSVQLPLVWNIQVSTWLRHYVYDRLVQNGKRPGFFQLLATQTVSAIWHGLYPGYIIFFVQSALMIAGSRVIYRWQQAVPPKMGLVKNIFVFFNFAYTLLVLNYSAVGFMVLSMHETLASYGSVYYIGTILPITLILLSYVIKPGKPARSKAHKEQ CpauLPCAT SEQ ID NO: 191MELEIGSVAAAIGVSVPVARFLLCFLATIPVSFLCRLLPARLPKHLYSAASGAILSYLSFGPSSNLHFIVPMSLGYLSMLFFRPFSGLLTFFLGFGYLIGCHVYYMSGDAWKEGGIDATGALMVLTLKVISCSINYNDGLLKEEGLRESQKKNRLTKMPSLIEYFGYCLCCGSHFAGPVYEMKDYLEWTEGKGIWSRSEKDPKPSPFGGALRAIIQAAVCMAMHMYLVPHHPLTRFTEPVYYEWGFFRRLSYQYMAAQTARWKYYFIWSISEASLIISGLGFSGWTESSPPKPRWDKAKNVDIIGVEFAKSSVQLPLVWNIQVSTWLRHYVYDRLVQNGKRPGFFQLLATQTVSAVWHGLYPGYIIFFVQSALMIAGSRVIYRWQQAVPQKMGLVKNIFVFFNFAYTLLVLNYSAVGFMVLSMHETLASYGSVYYIGTILPITLILLSYVIKPGKPTRSKVHKEQ CschuLPCAT SEQ ID NO: 192MELEMEPLAAAIGVSVAVFRFLVCFIATIPVSFICRLVPGGLPRHLFSAASGAVLSYLSFGFSSNLHFLVPMTLGYLSMILFRRFCGILTFFLGFGYLIGCHVYYMSGDAWKEGGIDATGALMVLTLKVISCSINYNDGLLKEEGLRESQKKNRLIRLPSLIEYFGYCLCCGSHFAGPVYEMKDYLDWTEGKGIWSHSEKGPKPSPLRAALRAIIQAGFCMAMYLYLVPHYPLTRFTDPVYYEWGILRRLSYQYMASFTARWKYYFIWSISEASLIISGLGFSGWTESSPPKPRWDRAKNVDILGVELAKSSVQIPLVWNIQVSTWLRHYVYDRLVQNGKRPGFLQLLATQTVSAIWHGVYPGYLIFFVQSALMIAGSRAIYRWQQAVPPKMSLVKNTLVFFNFAYTLLVLNYSAVGFMVLSMHETLASYGSVYYVGTILPVTLILLGYVIKPGKSPRSKASKEQ CawgPLA2-1 SEQ ID NO: 193MNFDFLSNIPWFGAKASDNAGSSFGSATIVIQQPPPVSRGFDIRHWGWPWSVLSVLPWGKPGCDELRAPPTTINRRLKRNATSMHSSAVRGNAEAARVRFRPYVSKVPWHTGFRGLLSQLFPRYGHYCGPNWSSGKNGGSPVWDQRPIDWLDYCCYCHDIGYDTHDQAKLLEADLAFLECLERPSYPTKGDAHVAHMYKTMCVTGLRNVLIPYRTQLLRLNSRQPLIDFGWLSNAAWKGWNAQKS CignPLA2-1 SEQ ID NO: 194MNLDFLSKI PWFEAKASENPGLNLGSTTIVIKQPRQGFDIRHWGWPWSVLTWGNRVTDEVHAPPTTINRRLKRNATGPAVQGDTEAARLRFRPYVSKVPWHTGFRGLLSQLFPRYGHYCGPNWSSGKNGGSPVWDQRPIDWLDYCCYCHDIGYDTHDQAKLLEADLAFLECLERPSYPTTGDAHVAHMYKTMCVTGLRNVLIPYRTQLLRLNFRQPLIDF GWLSNAAWKGWSAQKTCuPSR23PLA2-2 SEQ ID NO: 195MVHLPHTLKLGLVIAISISGLCFSSTPARALNVGIQAAGVTVSVGKGCSRKCESDFCKVPPFLRYGKYCGLMYSGCPGEKPCDGLDACCMKHDACVQAKNNDYLSQECSQNLLNCMASFRMSGGKQFKGSTCQVDEVVDVLTVVMEAALLAGRYLHKP CprocPLA2-2SEQ ID NO: 196 MVHLPHTLKLGLVIAISISGLCLSSTPARALNVGIQAAGVTVSVGKGCSRKCESDFCKVPPFLRYGKYCGLMYSGCPGEKPCDGLDACCMKHDACVQAKNDDYLSQECSQNLLNCMASFRMSGGKQFKGSTCQVDEVVDVLTVVMEAALLAGRYLHKP pSZ4329 SEQ ID NO: 197agcggaagagcgcccaatgtttaaacccctcaactgcgacgctgggaaccttctccgggcaggcgatgtgcgtgggtttgcctccttggcacggctctacaccgtcgagtacgccatgaggcggtgatggctgtgtcggttgccacttcgtccagagacggcaagtcgtccatcctctgcgtgtgtggcgcgacgctgcagcagtccctctgcagcagatgagcgtgactttggccatttcacgcactcgagtgtacacaatccatttttcttaaagcaaatgactgctgattgaccagatactgtaacgctgatttcgctccagatcgcacagatagcgaccatgttgctgcgtctgaaaatctggattccgaattcgaccctggcgctccatccatgcaacagatggcgacacttgttacaattcctgtcacccatcggcatggagcaggtccacttagattcccgatcacccacgcacatctcgctaatagtcattcgttcgtgtcttcgatcaatctcaagtgagtgtgcatggatcttggttgacgatgcggtatgggtttgcgccgctggctgcagggtctgcccaaggcaagctaacccagctcctctccccgacaatactctcgcaggcaaagccggtcacttgccttccagattgccaataaactcaattatggcctctgtcatgccatccatgggtctgatgaatggtcacgctcgtgtcctgaccgttccccagcctctggcgtcccctgccccgcccaccagcccacgccgcgcggcagtcgctgccaaggctgtctcggaggtaccctttcttgcgctatgacacttccagcaaaaggtagggcgggctgcgagacggcttcccggcgctgcatgcaacaccgatgatgcttcgaccccccgaagctccttcggggctgcatgggcgctccgatgccgctccagggcgagcgctgtttaaatagccaggcccccgattgcaaagacattatagcgagctaccaaagccatattcaaacacctagatcactaccacttctacacaggccactcgagcttgtgatcgcactccgctaagggggcgcctcttcctcttcgtttcagtcacaacccgcaaactctagaatatcaatgctgctgcaggccttcctgttcctgctggccggcttcgccgccaagatcagcgcctccatgacgaacgagacgtccgaccgccccctggtgcacttcacccccaacaagggctggatgaacgaccccaacggcctgtggtacgacgagaaggacgccaagtggcacctgtacttccagtacaacccgaacgacaccgtctgggggacgcccttgttctggggccacgccacgtccgacgacctgaccaactgggaggaccagcccatcgccatcgccccgaagcgcaacgactccggcgccttctccggctccatggtggtggactacaacaacacctccggcttcttcaacgacaccatcgacccgcgccagcgctgcgtggccatctggacctacaacaccccggagtccgaggagcagtacatctcctacagcctggacggcggctacaccttcaccgagtaccagaagaaccccgtgctggccgccaactccacccagttccgcgacccgaaggtcttctggtacgagccctcccagaagtggatcatgaccgcggccaagtcccaggactacaagatcgagatctactcctccgacgacctgaagtcctggaagctggagtccgcgttcgccaacgagggcttcctcggctaccagtacgagtgccccggcctgatcgaggtccccaccgagcaggaccccagcaagtcctactgggtgatgttcatctccatcaaccccggcgccccggccggcggctccttcaaccagtacttcgtcggcagcttcaacggcacccacttcgaggccttcgacaaccagtcccgcgtggtggacttcggcaaggactactacgccctgcagaccttcttcaacaccgacccgacctacgggagcgccctgggcatcgcgtgggcctccaactgggagtactccgccttcgtgcccaccaacccctggcgctcctccatgtccctcgtgcgcaagttctccctcaacaccgagtaccaggccaacccggagacggagctgatcaacctgaaggccgagccgatcctgaacatcagcaacgccggcccctggagccggttcgccaccaacaccacgttgacgaaggccaacagctacaacgtcgacctgtccaacagcaccggcaccctggagttcgagctggtgtacgccgtcaacaccacccagacgatctccaagtccgtgttcgcggacctctccctctggttcaagggcctggaggaccccgaggagtacctccgcatgggcttcgaggtgtccgcgtcctccttcttcctggaccgcgggaacagcaaggtgaagttcgtgaaggagaacccctacttcaccaaccgcatgagcgtgaacaaccagcccttcaagagcgagaacgacctgtcctactacaaggtgtacggcttgctggaccagaacatcctggagctgtacttcaacgacggcgacgtcgtgtccaccaacacctacttcatgaccaccgggaacgccctgggctccgtgaacatgacgacgggggtggacaacctgttctacatcgacaagttccaggtgcgcgaggtcaagtgacaattgacgcccgcgcggcgcacctgacctgttctctcgagggcgcctgttctgccttgcgaaacaagcccctggagcatgcgtgcatgatcgtctctggcgccccgccgcgcggtttgtcgccctcgcgggcgccgcggccgcgggggcgcattgaaattgttgcaaaccccacctgacagattgagggcccaggcaggaaggcgttgagatggaggtacaggagtcaagtaactgaaagtttttatgataactaacaacaaagggtcgtttctggccagcgaatgacaagaacaagattccacatttccgtgtagaggcttgccatcgaatgtgagcgggcgggccgcggacccgacaaaacccttacgacgtggtaagaaaaacgtggcgggcactgtccctgtagcctgaagaccagcaggagacgatcggaagcatcacagcacaggatcccgcgtctcgaacagagcgcgcagaggaacgctgaaggtctcgcctctgtcgcacctcagcgcggcatacaccacaataaccacctgacgaatgcgcttggttcttcgtccattagcgaagcgtccggttcacacacgtgccacgttggcgaggtggcaggtgacaatgatcggtggagctgatggtcgaaacgttca cagcctagggatatcgcctgctcaagcgggcgctcaacatgcagagcgtcagcgagacgggctgtggcgatcgcgagacggacgaggccgcctctgccctgtttgaactgagcgtcagcgctggctaaggggagggagactcatccccaggctcgcgccagggctctgatcccgtctcgggcggtgatcggcgcgcatgactacgacccaacgacgtacgagactgatgtcggtcccgacgaggagcgccgcgaggcactcccgggccaccgaccatgtttacaccgaccgaaagcactcgctcgtatccattccgtgcgcccgcacatgcatcatcttttggtaccgacttcggtcttgttttacccctacgacctgccttccaaggtgtgagcaactcgcccggacatgaccgagggtgatcatccggatccccaggccccagcagcccctgccagaatggctcgcgctttccagcctgcaggcccgtctcccaggtcgacgcaacctacatgaccaccccaatctgtcccagaccccaaacaccctccttccctgcttctctgtgatcgctgatcagcaacaactagtaacaatggccaccgcctccaccttctccgccttcaacgcccgctgcggcgacctgcgccgctccgccggctccggcccccgccgccccgcccgccccctgcccgtgcgcgccgccatcaacgactccgcccaccccaaggccaacggctccgccgtgagcctgaagagcggcagcctgaacacccaggaggacacctcctccagcccccccccccgcaccttcctgcaccagctgcccgactggagccgcctgctgaccgccatcaccaccgtgttcgtgaagtccaagcgccccgacatgcacgaccgcaagtccaagcgccccgacatgctggtggacagcttcggcctggagtccaccgtgcaggacggcctggtgttccgccagtccttctccatccgctcctacgagatcggcaccgaccgcaccgccagcatcgagaccctgatgaaccacctgcaggagacctccctgaaccactgcaagagcaccggcatcctgctggacggcttcggccgcaccctggagatgtgcaagcgcgacctgatctgggtggtgatcaagatgcagatcaaggtgaaccgctaccccgcctggggcgacaccgtggagatcaacacccgcttcagccgcctgggcaagatcggcatgggccgcgactggctgatctccgactgcaacaccggcgagatcctggtgcgcgccaccagcgcctacgccatgatgaaccagaagacccgccgcctgtccaagctgccctacgaggtgcaccaggagatcgtgcccctgttcgtggacagccccgtgatcgaggactccgacctgaaggtgcacaagttcaaggtgaagaccggcgacagcatccagaagggcctgacccccggctggaacgacctggacgtgaaccagcacgtgtccaacgtgaagtacatcggctggatcctggagagcatgcccaccgaggtgctggagacccaggagctgtgctccctggccctggagtaccgccgcgagtgcggccgcgactccgtgctggagagcgtgaccgccatggaccccagcaaggtgggcgtgcgctcccagtaccagcacctgctgcgcctggaggacggcaccgccatcgtgaacggcgccaccgagtggcgccccaagaacgccggcgccaacggcgccatctccaccggcaagaccagcaacggcaactccgtgtccatggactacaaggaccacgacggcgactacaaggaccacgacatcgactacaaggacgacgacgacaagtgactcgaggcagcagcagctcggatagtatcgacacactctggacgctggtcgtgtgatggactgttgccgccacacttgctgccttgacctgtgaatatccctgccgcttttatcaaacagcctcagtgtgtttgatcttgtgtgtacgcgcttttgcgagttgctagctgcttgtgctatttgcgaataccacccccagcatccccttccctcgtttcatatcgcttgcatcccaaccgcaacttatttacgctgtcctgctatccctcagcgctgctcctgctcctgctcactgccctcgcacagccttggtttgggctccgcctgtattctcctggtactgcaacctgtaaaccagcactgcaatgctgatgcacgggaagtagtgggatgggaacacaaatggaaagcttgagctccagcgccatgccacgccctttgatggcttcaagtacgattacggtgttggattgtgtgtttgttgcgtagtgtgcatggtttagaataatacacttgatttcttgctcacggcaatctcggcttgtccgcaggttcaaccccatttcggagtctcaggtcagccgcgcaatgaccagccgctacttcaaggacttgcacgacaacgccgaggtgagctatgtttaggacttgattggaaattgtcgtcgacgcatattcgcgctccgcgacagcacccaagcaaaatgtcaagtgcgttccgatttgcgtccgcaggtcgatgttgtgatcgtcggcgccggatccgccggtctgtcctgcgcttacgagctgaccaagcaccctgacgtccgggtacgcgagctgagattcgattagacataaattgaagattaaacccgtagaaaaatttgatggtcgcgaaactgtgctcgattgcaagaaattgatcgtcctccactccgcaggtcgccatcatcgagcagggcgttgctcccggcggcggcgcctggctggggggacagctgttctcggccatgtgtgtacgtagaaggatgaatttcagctggttttcgttgcacagctgtttgtgcatgatttgtttcagactattgttgaatgtttttagatttcttaggatgcatgatttgtctgcatgcgactgaagagcgttt pSZ5078 SEQ ID NO: 198agcggaagagcgcccaatgtttaaacccctcaactgcgacgctgggaaccttctccgggcaggcgatgtgcgtgggtttgcctccttggcacggctctacaccgtcgagtacgccatgaggcggtgatggctgtgtcggttgccacttcgtccagagacggcaagtcgtccatcctctgcgtgtgtggcgcgacgctgcagcagtccctctgcagcagatgagcgtgactttggccatttcacgcactcgagtgtacacaatccatttttcttaaagcaaatgactgctgattgaccagatactgtaacgctgatttcgctccagatcgcacagatagcgaccatgttgctgcgtctgaaaatctggattccgaattcgaccctggcgctccatccatgcaacagatggcgacacttgttacaattcctgtcacccatcggcatggagcaggtccacttagattcccgatcacccacgcacatctcgctaatagtcattcgttcgtgtcttcgatcaatctcaagtgagtgtgcatggatcttggttgacgatgcggtatgggtttgcgccgctggctgcagggtctgcccaaggcaagctaacccagctcctctccccgacaatactctcgcaggcaaagccggtcacttgccttccagattgccaataaactcaattatggcctctgtcatgccatccatgggtctgatgaatggtcacgctcgtgtcctgaccgttccccagcctctggcgtcccctgccccgcccaccagcccacgccgcgcggcagtcgctgccaaggctgtctcggaggtaccctttcttgcgctatgacacttccagcaaaaggtagggcgggctgcgagacggcttcccggcgctgcatgcaacaccgatgatgcttcgaccccccgaagctccttcggggctgcatgggcgctccgatgccgctccagggcgagcgctgtttaaatagccaggcccccgattgcaaagacattatagcgagctaccaaagccatattcaaacacctagatcactaccacttctacacaggccactcgagcttgtgatcgcactccgctaagggggcgcctcttcctcttcgtttcagtcacaacccgcaaactctagaatatcaatgctgctgcaggccttcctgttcctgctggccggcttcgccgccaagatcagcgcctccatgacgaacgagacgtccgaccgccccctggtgcacttcacccccaacaagggctggatgaacgaccccaacggcctgtggtacgacgagaaggacgccaagtggcacctgtacttccagtacaacccgaacgacaccgtctgggggacgcccttgttctggggccacgccacgtccgacgacctgaccaactgggaggaccagcccatcgccatcgccccgaagcgcaacgactccggcgccttctccggctccatggtggtggactacaacaacacctccggcttcttcaacgacaccatcgacccgcgccagcgctgcgtggccatctggacctacaacaccccggagtccgaggagcagtacatctcctacagcctggacggcggctacaccttcaccgagtaccagaagaaccccgtgctggccgccaactccacccagttccgcgacccgaaggtcttctggtacgagccctcccagaagtggatcatgaccgcggccaagtcccaggactacaagatcgagatctactcctccgacgacctgaagtcctggaagctggagtccgcgttcgccaacgagggcttcctcggctaccagtacgagtgccccggcctgatcgaggtccccaccgagcaggaccccagcaagtcctactgggtgatgttcatctccatcaaccccggcgccccggccggcggctccttcaaccagtacttcgtcggcagcttcaacggcacccacttcgaggccttcgacaaccagtcccgcgtggtggacttcggcaaggactactacgccctgcagaccttcttcaacaccgacccgacctacgggagcgccctgggcatcgcgtgggcctccaactgggagtactccgccttcgtgcccaccaacccctggcgctcctccatgtccctcgtgcgcaagttctccctcaacaccgagtaccaggccaacccggagacggagctgatcaacctgaaggccgagccgatcctgaacatcagcaacgccggcccctggagccggttcgccaccaacaccacgttgacgaaggccaacagctacaacgtcgacctgtccaacagcaccggcaccctggagttcgagctggtgtacgccgtcaacaccacccagacgatctccaagtccgtgttcgcggacctctccctctggttcaagggcctggaggaccccgaggagtacctccgcatgggcttcgaggtgtccgcgtcctccttcttcctggaccgcgggaacagcaaggtgaagttcgtgaaggagaacccctacttcaccaaccgcatgagcgtgaacaaccagcccttcaagagcgagaacgacctgtcctactacaaggtgtacggcttgctggaccagaacatcctggagctgtacttcaacgacggcgacgtcgtgtccaccaacacctacttcatgaccaccgggaacgccctgggctccgtgaacatgacgacgggggtggacaacctgttctacatcgacaagttccaggtgcgcgaggtcaagtgacaattgacgcccgcgcggcgcacctgacctgttctctcgagggcgcctgttctgccttgcgaaacaagcccctggagcatgcgtgcatgatcgtctctggcgccccgccgcgcggtttgtcgccctcgcgggcgccgcggccgcgggggcgcattgaaattgttgcaaaccccacctgacagattgagggcccaggcaggaaggcgttgagatggaggtacaggagtcaagtaactgaaagtttttatgataactaacaacaaagggtcgtttctggccagcgaatgacaagaacaagattccacatttccgtgtagaggcttgccatcgaatgtgagcgggcgggccgcggacccgacaaaacccttacgacgtggtaagaaaaacgtggcgggcactgtccctgtagcctgaagaccagcaggagacgatcggaagcatcacagcacaggatcccgcgtctcgaacagagcgcgcagaggaacgctgaaggtctcgcctctgtcgcacctcagcgcggcatacaccacaataaccacctgacgaatgcgcttggttcttcgtccattagcgaagcgtccggttcacacacgtgccacgttggcgaggtggcaggtgacaatgatcggtggagctgatggtcgaaacgttcacagcctagggatatcgaattcggccgacaggacgcgcgtcaaaggtgctggtcgtgtatgccctggccggcaggtcgttgctgctgctggttagtgattccgcaaccctgattttggcgtcttattttggcgtggcaaacgctggcgcccgcgagccgggccggcggcgatgcggtgccccacggctgccggaatccaagggaggcaagagcgcccgggtcagttgaagggctttacgcgcaaggtacagccgctcctgcaaggctgcgtggtggaattggacgtgcaggtcctgctgaagttcctccaccgcctcaccagcggacaaagcaccggtgtatcaggtccgtgtcatccactctaaagagctcgactacgacctactgatggccctagattcttcatcaaaaacgcctgagacacttgcccaggattgaaactccctgaagggaccaccaggggccctgagttgttccttccccccgtggcgagctgccagccaggctgtacctgtgatcgaggctggcgggaaaataggcttcgtgtgctcaggtcatgggaggtgcaggacagctcatgaaacgccaacaatcgcacaattcatgtcaagctaatcagctatttcctcttcacgagctgtaattgtcccaaaattctggtctaccgggggtgatccttcgtgtacgggcccttccctcaaccctaggtatgcgcgcatgcggtcgccgcgcaa ctcgcgcgagggccgagggtttgggacgggccgtcccgaaatgcagttgcacccggatgcgtggcacctiftttgcgataatttatgcaatggactgctctgcaaaattctggctctgtcgccaaccctaggatcagcggcgtaggatttcgtaatcattcgtcctgatggggagctaccgactaccctaatatcagcccgactgcctgacgccagcgtccacttttgtgcacacattccattcgtgcccaagacatttcattgtggtgcgaagcgtccccagttacgctcacctgtttcccgacctccttactgttctgtcgacagagcgggcccacaggccggtcgcagccactagtatggccaccgcctccaccttctccgccttcaacgcccgctgcggcgacctgcgccgctccgccggctccggcccccgccgccccgcccgccccctgcccgtgcgcgccgccatcaactcccgcgcccaccccaaggccaacggctccgccgtgtccctgaagtccggctccctgaacacccaggaggacacctcctcctccccccccccccgcaccttcctgcaccagctgcccgactggtcccgcctgctgaccgccatcaccaccgtgttcgtgaagtccaagcgccccgacatgcacgaccgcaagtccaagcgccccgacatgctgatggactccttcggcctggagtccatcgtgcaggagggcctggagttccgccagtccttctccatccgctcctacgagatcggcaccgaccgcaccgcctccatcgagaccctgatgaactacctgcaggagacctccctgaaccactgcaagtccaccggcatcctgctggacggcttcggccgcacccccgagatgtgcaagcgcgacctgatctgggtggtgaccaagatgaagatcaaggtgaaccgctaccccgcctggggcgacaccgtggagatcaacacctggttctcccgcctgggcaagatcggcaagggccgcgactggctgatctccgactgcaacaccggcgagatcctgatccgcgccacctccgcctacgccaccatgaaccagaagacccgccgcctgtccaagctgccctacgaggtgcaccaggagatcgcccccctgttcgtggactccccccccgtgatcgaggacaacgacctgaagctgcacaagttcgaggtgaagaccggcgactccatccacaagggcctgacccccggctggaacgacctggacgtgaaccagcacgtgtccaacgtgaagtacatcggctggatcctggagtccatgcccaccgaggtgctggagacccaggagctgtgctccctggccctggagtaccgccgcgagtgcggccgcgactccgtgctggagtccgtgaccgccatggaccccaccaaggtgggcggccgctcccagtaccagcacctgctgcgcctggaggacggcaccgacatcgtgaagtgccgcaccgagtggcgccccaagaaccccggcgccaacggcgccatctccaccggcaagacctccaacggcaactccgtgtccatggactacaaggaccacgacggcgactacaaggaccacgacatcgactacaaggacgacgacgacaagtgattaattaactcgaggcagcagcagctcggatagtatcgacacactctggacgctggtcgtgtgatggactgttgccgccacacttgctgccttgacctgtgaatatccctgccgcttttatcaaacagcctcagtgtgtttgatcttgtgtgtacgcgcttttgcgagttgctagctgcttgtgctatttgcgaataccacccccagcatccccttccctcgtttcatatcgcttgcatcccaaccgcaacttatctacgctgtcctgctatccctcagcgctgctcctgctcctgctcactgcccctcgcacagccttggtttgggctccgcctgtattctcctggtactgcaacctgtaaaccagcactgcaatgctgatgcacgggaagtagtgggatgggaacacaaatggaaagcttgagctccagcgccatgccacgccctttgatggcttcaagtacgattacggtgttggattgtgtgtttgttgcgtagtgtgcatggtttagaataatacacttgatttcttgctcacggcaatctcggcttgtccgcaggttcaaccccatttcggagtctcaggtcagccgcgcaatgaccagccgctacttcaaggacttgcacgacaacgccgaggtgagctatgtttaggacttgattggaaattgtcgtcgacgcatattcgcgctccgcgacagcacccaagcaaaatgtcaagtgcgttccgatttgcgtccgcaggtcgatgttgtgatcgtcggcgccggatccgccggtctgtcctgcgcttacgagctgaccaagcaccctgacgtccgggtacgcgagctgagattcgattagacataaattgaagattaaacccgtagaaaaatttgatggtcgcgaaactgtgctcgattgcaagaaattgatcgtcctccactccgcaggtcgccatcatcgagcagggcgttgctcccggcggcggcgcctggctggggggacagctgttctcggccatgtgtgtacgtagaaggatgaatttcagctggttttcgttgcacagctgtttgtgcatgatttgtttcagactattgttgaatgtttttagatttcttaggatgcatgatttgtctgcatgcgactgaagcgtttaaaccgcct

What is claimed:
 1. A microalgal cell having an ablated or downregulatedfatty acyl-ACP thioesterase (FATA) gene, wherein the cell is modified toexpress a heterologous lysophosphatidic acid acyltransferase (LPAAT)comprising an amino acid sequence that has at least 80% identity to anacyltransferase encoded by SEQ ID NO: 90, 89, 92, 93 or 95 and whereinthe modified microalgal cell produces an oil with an elevated ratio ofsaturated-unsaturated-saturated triglycerides over trisaturatedtriglycerides as compared to a corresponding unmodified cell.
 2. Themicroalgal cell of claim 1, wherein the cell is modified to coexpresswith the heterologous LPAAT at least one exogenous gene that encodes anenzyme selected from the group consisting of invertase, a fatty acyl-ACPthioesterase, a melibiase, a ketoacyl synthase and a THIC.
 3. Themicroalgal cell of claim 1, wherein the cell is modified to ablate ordownregulate the expression of at least one endogenous gene selectedfrom the group consisting of: a stearoyl ACP desaturase, a fatty acyldesaturase, a fatty acyl-ACP thioesterase (FATA or FATB), a ketoacylsynthase (KASI, KASII, KASIII or KAS IV) and an acyltransferase (DGAT,GPAT or LPCAT).
 4. The microalgal cell of claim 2, wherein the cell isfurther modified to overexpress a gene encoding a C18:0-specific FATA1thioesterase.
 5. The microalgal cell of claim 4, wherein theC18:0-specific FATA1 thioesterase is a variant Garcinia thioesterase. 6.The microalgal cell of claim 5, wherein the variant Garciniathioesterase has at least 80% identity to SEQ ID NO:
 142. 7. Themicroalgal cell of claim 6, wherein the variant Garcinia thioesterasecomprises one or more of amino acid variants selected from the groupconsisting of L91F, L91K, L91S, G96A, G96T, G96V, G108A, G108V, S111A,S111V T156F, T156A, T156K, T156V and V193A.
 8. The microalgal cell ofclaim 7, wherein the variant Garcinia thioesterase is a variantcomprising the substitutions S111A and V193A, a variant comprising thesubstitution G96A, or a variant comprising the substitution G108A. 9.The microalgal cell of claim 2, wherein the ketoacyl synthase is aKASII.
 10. The microalgal cell of claim 4, wherein the ketoacyl synthaseis a KASII.
 11. The microalgal cell of claim 3, wherein the cell ismodified to ablate or downregulate the expression of an endogenousstearoyl ACP desaturase-2 (SAD2) gene and an endogenous fatty acyldesaturase-2 (FAD2) gene.
 12. The microalgal cell of claim 10, whereinthe cell is modified to ablate or downregulate the expression of anendogenous stearoyl ACP desaturase-2 (SAD2) gene and an endogenous fattyacyl desaturase-2 (FAD2) gene.
 13. The microalgal cell of claim 1,wherein the cell is modified to express a Theobroma cacao diacylglycerolO-acyltransferase.
 14. The microalgal cell of claim 13, wherein theTheobroma cacao diacylglycerol O-acyltransferase is a Theobroma cacaodiacylglycerol O-acyltransferase-1 or a Theobroma cacao diacylglycerolO-acyltransferase-2.
 15. The microalgal cell of claim 12, wherein theTheobroma cacao diacylglycerol O-acyltransferase is a Theobroma cacaodiacylglycerol O-acyltransferase-1 or a Theobroma cacao diacylglycerolO-acyltransferase-2.
 16. The microalgal cell of claim 1, wherein thecell is of the genus Prototheca or Chlorella.
 17. The microalgal cell ofclaim 16, wherein the cell is a Prototheca moriformis cell.
 18. Themicroalgal cell of claim 15, wherein the cell is a Prototheca moriformiscell.
 19. A method of producing an oil comprising: (a) cultivating themicroalgal cell of claim 1 under conditions to produce the oil; and (b)extracting the oil from the microalgal cell; wherein the oil comprisesat least 50% stearate-oleate-stearate (SOS) triglycerides with anelevated ratio of saturated-unsaturated-saturated triglycerides overtrisaturated triglycerides as compared to a corresponding unmodifiedcell.
 20. A method of producing an oil comprising: (a) cultivating themicroalgal cell of claim under conditions to produce the oil; and (b)extracting the oil from the microalgal cell; wherein the oil comprisesat least 60% stearate-oleate-stearate (SOS) triglycerides, less than 5%trisaturates and wherein the fatty acid profile of the oil comprises atleast 50% C18:0.